Methods of treating cerebral edema

ABSTRACT

The present invention relates to the use of selective aquaporin inhibitors, e.g., of aquaporin-4 or aquaporin-2, e.g., certain phenylbenzamide compounds, for the prophylaxis, treatment and control of aquaporin-mediated conditions, e.g., diseases of water imbalance, for example edema (particularly edema of the brain and spinal cord, e.g., following trauma or ischemic stroke, as well as the edema associated with glioma, meningitis, acute mountain sickness, epileptic seizures, infections, metabolic disorders, hypoxia, water intoxication, hepatic failure, hepatic encephalopathy, diabetic ketoacidosis, abscess, eclampsia, Creutzfeldt-Jakob disease, and lupus cerebritis, as well as edema consequent to microgravity and/or radiation exposure, as well as edema consequent to invasive central nervous system procedures, e.g., neurosurgery, endovascular clot removal, spinal tap, aneurysm repair, or deep brain stimulation, as well as retinal edema), as well as hyponatremia and excess fluid retention, and diseases such as epilepsy, retinal ischemia and other diseases of the eye associated with abnormalities in intraocular pressure and/or tissue hydration, myocardial ischemia, myocardial ischemia/reperfusion injury, myocardial infarction, myocardial hypoxia, congestive heart failure, sepsis, and neuromyelitis optica, as well as migraines, as well as to novel assays for identifying aquaporin inhibitors.

The subject matter of this case is related to U.S. ProvisionalApplication 61/644,268, filed May 8, 2012, U.S. Provisional Application61/651,778, filed May 25, 2012, and U.S. Provisional Application61/799,606, filed Mar. 15, 2013, the contents of each of which areincorporated herein by reference.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under Grant Numbers R43NS060199 and R44 NS060199 awarded by the National Institutes of Health.The government has certain rights in the invention.

FIELD OF THE INVENTION

The present invention relates to the use of selective aquaporininhibitors, e.g., of aquaporin-4 or aquaporin-2, e.g., certainphenylbenzamide compounds, for the prophylaxis, treatment and control ofaquaporin-mediated conditions, e.g., diseases of water imbalance, forexample edema (particularly edema of the brain and spinal cord, e.g.,following trauma or ischemic stroke, as well as the edema associatedwith glioma, meningitis, acute mountain sickness, epileptic seizures,infections, metabolic disorders, hypoxia, water intoxication, hepaticfailure, hepatic encephalopathy, diabetic ketoacidosis, abscess,eclampsia, Creutzfeldt-Jakob disease, and lupus cerebritis, as well asedema consequent to microgravity and/or radiation exposure, as well asedema consequent to invasive central nervous system procedures, e.g.,neurosurgery, endovascular clot removal, spinal tap, aneurysm repair, ordeep brain stimulation, as well as retinal edema), as well ashyponatremia and excess fluid retention, and diseases such as epilepsy,retinal ischemia and other diseases of the eye associated withabnormalities in intraocular pressure and/or tissue hydration,myocardial ischemia, myocardial ischemia/reperfusion injury, myocardialinfarction, myocardial hypoxia, congestive heart failure, sepsis, andneuromyelitis optica, as well as migraines, as well as to novel assaysfor identifying aquaporin inhibitors.

BACKGROUND OF THE INVENTION

Aquaporins are cell membrane proteins that act as molecular waterchannels to mediate the flow of water in and out of the cells. Whilethere is some degree of passive diffusion or osmosis of water acrosscell membranes, the rapid and selective transport of water in and out ofcells involves aquaporins. These water channels selectively conductwater molecules in and out of the cell, while blocking the passage ofions and other solutes, thereby preserving the membrane potential of thecell. Aquaporins are found in virtually all life forms, from bacteria toplants to animals. In humans, they are found in cells throughout thebody.

Cerebral edema (CE) is a major contributor to stroke damage, as it canresult in increased intracerebral pressure (ICP), a correspondingdecrease in cerebral perfusion, and potentially permanent or fatal braindamage. Edema also contributes to CNS damage in, for example, traumaticbrain and spinal cord injury, glioma, meningitis, acute mountainsickness, epileptic seizures, infections, metabolic disorders, hypoxia,water intoxication, hepatic failure, hepatic encephalopathy, diabeticketoacidosis, abscess, eclampsia, Creutzfeldt-Jakob disease, and lupuscerebritis. Patients surviving the period of maximal ICP, usually thethree days following a stroke or traumatic brain injury, are likely tosurvive. Unfortunately, only a few treatment options are available forCE, and these are of limited efficacy.

Hyponatremia, characterized by serum sodium levels ≦135 mM, is the mostcommon form of electrolyte imbalance with hospitals nationwide reportingan incidence of 15-20%. The associated fluid retention is symptomatic ofheart failure (HF), liver cirrhosis, nephrotic disorder, and syndrome ofinappropriate antidiuretic hormone secretion (SIADH). Various diureticsare used to treat congestion associated with HF. By inhibiting theNa/K/Cl cotransporter in the thick ascending loop of Henle, loopdiuretics cause natriuresis by decreasing Na⁺ and Cl⁻ reabsorption fromthe urine. An alternative therapy for hyponatremia is the use ofvasopressin receptor antagonists, which inhibit water reabsorption byinhibiting the vasopressin-induced trafficking of AQP2. Unfortunately,both loop diuretics and vasopressin receptor antagonists act indirectlytoward a desired physiological outcome. An ideal drug would block waterreabsorption directly, thus minimizing potential side-effects caused byupstream effectors, but no such drugs are currently known.

Epilepsy is a brain disorder characterized by recurrent seizures.Seizures occur because of disturbed brain activity resulting in somedegree of temporary brain dysfunction. Seizures may cause uncontrollableshaking and loss of consciousness but, more commonly, a personexperiencing a seizure stops moving or becomes unaware of what ishappening Anticonvulsants may be used to treat epilepsy, howeveranticonvulsants are not effective for all people with epilepsy.

Ischemia is a condition characterized by an interruption or inadequatesupply of blood to tissues. Retinal ischemia occurs due to a deficientsupply of blood to the retina. Vascular occlusion, glaucoma, anddiabetic retinopathy are associated with retinal ischemia and canproduce retinal edema and ganglion cell death leading to visualimpairment and blindness. AQP4 is expressed in the Müller cells in theretina. Due to relatively ineffectual treatment, retinal ischemiaremains a common cause of visual impairment and blindness.

Myocardial ischemia is a condition caused by a blockage or constrictionof one more of the coronary arteries, such as can occur withatherosclerotic plaque occlusion or rupture. Myocardial infarction, aheart attack, occurs when myocardial ischemia exceeds a criticalthreshold and overwhelms myocardial cellular repair mechanisms designedto maintain normal operating function and homeostasis. Myocardialinfarction remains a leading cause of morbidity and mortality worldwide.Compounds effective in treating myocardial ischemia, myocardialischemia/reperfusion injury, myocardial infarction, and congestive heartfailure would be useful pharmaceuticals.

Phenylbenzamide compounds are known as pharmaceuticals. Phenylbenzamidesinclude compounds such niclosamide(5-chloro-N-(2-chloro-4-nitrophenyl)-2-hydroxybenzamide), anantihemintic agent used to treat tapeworms, but are not known to haveany effect on aquaporins. US Patent Publication US 2010/0274051 A1 (thecontents of which are incorporated herein by reference) describe certainphenylbenzamides as being useful to inhibit NF—KB via selectiveinhibition of IKK-β, while U.S. Pat. No. 7,626,042 (also incorporatedherein by reference) discloses O-acyl derivatives of such compounds,while U.S. Pat. No. 7,700,655 (also incorporated herein by reference)describe certain phenylbenzamides as being useful to treat allergicdiseases. These patent applications, however, do not disclose anythingabout cerebral edema or water imbalance (aquaresis) or aquaporins.

In a 2004 paper, a group purportedly investigated the efficacy ofN-(3,5-Bis-trifluoromethyl-phenyl)-5-chloro-2-hydroxy-benzamide in IκBphosphorylation blockade in a rat myocardial ischemia/reperfusion injurymodel. Onai, Y. et al., “Inhibition of IκB Phosphorylation inCardiomyocytes Attenuates Myocardial Ischemia/Reperfusion Injury,”Cardiovascular Research, 2004, 63, 51-59. The group reported someactivity. However, subsequently, the activity effect could not beconfirmed and accordinglyN-(3,5-Bis-trifluoromethyl-phenyl)-5-chloro-2-hydroxy-benzamide was notpursued for this indication in humans.

Prior to this invention, there are have been no known specific,validated inhibitors of aquaporins, for example AQP4 or AQP2. Certainantiepileptic or sulfonamide drugs (e.g., acetylsulfanilamide,acetazolamide, 6-ethoxy-benzothiazole-2-sulfonamide, topiramate,zonisamide, phenytoin, lamotrigine, and sumatriptan) were at one pointreported to be possible inhibitors of AQP4, but this later proved to beincorrect. Yang, et al., Bioorganic & Medicinal Chemistry (2008) 16:7489-7493. No direct inhibitors of AQP2 have been reported. The searchfor therapeutically useful aquaporin inhibitors has been hampered by alack of effective high throughput screening assays, as well as by a lackof highly selective inhibitors to develop and validate the assays and toserve as positive controls or binding competitors.

There is a great need for improved approaches to treating andcontrolling diseases of water imbalance, such as edema, for examplecerebral edema, and water retention and hyponatremia, as well asdiseases such as epilepsy, retinal ischemia, myocardial ischemia,myocardial ischemia/reperfusion injury, myocardial infarction,myocardial hypoxia, congestive heart failure, sepsis, and neuromyelitisoptica, as well as migraines.

BRIEF SUMMARY OF THE INVENTION

The invention provides the use of selective aquaporin inhibitors, e.g.,of aquaporin-4 or aquaporin-2 for the prophylaxis, treatment and controlof aquaporin-mediated conditions, e.g., diseases of water imbalance, forexample edema (particularly edema of the brain and spinal cord, e.g.,following trauma or ischemic stroke, as well as the edema associatedwith glioma, meningitis, acute mountain sickness, epileptic seizures,infections, metabolic disorders, water intoxication, hepatic failure,hepatic encephalopathy, diabetic ketoacidosis, abscess, eclampsia,Creutzfeldt-Jakob disease, and lupus cerebritis, as well as the edemaconsequent to microgravity and/or radiation exposure, as well as edemaconsequent to invasive central nervous system procedures, e.g.,neurosurgery, endovascular clot removal, spinal tap, aneurysm repair, ordeep brain stimulation, as well as retinal edema, as well as brainswelling consequent to cardiac arrest, e.g., related to the developmentof the metabolic acidosis (e.g. lactic acidosis) due to hypoxia beforethe resuscitation period), as well as hyponatremia and excess fluidretention, as well as diseases such as epilepsy, retinal ischemia andother diseases of the eye associated with abnormalities in intraocularpressure or tissue hydration, myocardial ischemia, myocardialischemia/reperfusion injury, myocardial infarction, myocardial hypoxia,congestive heart failure, sepsis, and neuromyelitis optica, as well asmigraines.

The invention further provides the use of certain phenylbenzamides toinhibit aquaporins, particularly AQP4 and AQP2.

The invention provides, inter alia, methods of treating or controlling adisease or condition mediated by an aquaporin, e.g., diseases orconditions of water imbalance and other diseases, for example,

-   -   edema of the brain or spinal cord, e.g., cerebral edema, e.g.        cerebral edema consequent to head trauma, ischemic stroke,        glioma, meningitis, acute mountain sickness, epileptic seizures,        infections, metabolic disorders, hypoxia (including general        systemic hypoxia and hypoxia due to cardiac arrest), water        intoxication, hepatic failure, hepatic encephalopathy, diabetic        ketoacidosis, abscess, eclampsia, Creutzfeldt-Jakob disease,        lupus cerebritis, or invasive central nervous system procedures,        e.g., neurosurgery, endovascular clot removal, spinal tap,        aneurysm repair, or deep brain stimulation or, e.g., spinal cord        edema consequent to spinal cord trauma, e.g., spinal cord        compression; or    -   cerebral and/or optical nerve edema consequent to microgravity        and/or radiation exposure; or    -   retinal edema; or    -   hyponatremia or excessive fluid retention, e.g., consequent to        heart failure (HF), liver cirrhosis, nephrotic disorder, or        syndrome of inappropriate antidiuretic hormone secretion        (SIADH); or    -   epilepsy, retinal ischemia or other diseases of the eye        associated with abnormalities in intraocular pressure and/or        tissue hydration, myocardial ischemia, myocardial        ischemia/reperfusion injury, myocardial infarction, myocardial        hypoxia, congestive heart failure, sepsis, or neuromyelitis        optica;    -   or migraines,        comprising administering to a patient in need thereof an        effective amount of an aquaporin inhibitor, e.g., an inhibitor        of AQP2 or AQP4, for example a phenylbenzamide, e.g.,        niclosamide or a compound of formula (I) as described in US        2010/0274051 A1 or U.S. Pat. No. 7,700,655, e.g., a compound of        general formulae (I), (I-1), (I-2), (I-3), and (I-4) as set        forth therein, e.g., selected from Compound Nos. 1-223 as set        forth in US 2010/0274051 or Compound Nos. 301-555 as set forth        in U.S. Pat. No. 7,700,655, or a compound of formula (I) as        described in U.S. Pat. No. 7,626,042, e.g., selected from        Compound Nos. 1-151 as set forth therein; for example a compound        of formula 1a:

-   -   wherein R₁, R₂, R₃, R₄, and R₅ are selected from H, halo,        halogenated C₁₋₄ alkyl (e.g., trifluoromethyl), and cyano; and        R₆ is selected from H and physiologically hydrolysable and        acceptable acyl groups,    -   in free or pharmaceutically acceptable salt form.

The invention further provides high throughput assays for identificationof specific aquaporins, comprising measuring the response of anaquaporin-expressing cell population versus a control cell population toa hypertonic or hypotonic solution in the presence or absence of a testcompound.

The invention further provides a compound of formula

Further areas of applicability of the present invention will becomeapparent from the detailed description provided hereinafter. It shouldbe understood that the detailed description and specific examples, whileindicating the preferred embodiment of the invention, are intended forpurposes of illustration only and are not intended to limit the scope ofthe invention.

BRIEF DESCRIPTION OF THE DRAWINGS

The present invention will become more fully understood from thedetailed description and the accompanying drawings, wherein:

FIG. 1 depicts results of aquaporin-4 (FIG. 1A) and aquaporin-2 (FIG.1B) mediated cell volume change assay, and the inhibitory effect ofCompound 3 (compound of formula 1a where R₁, R₃ and R₅ are each chloro,and R₂, R₄ and R₆ are H) against these aquaporins.

FIG. 2 depicts specificity of Compound 3 towards AQP-1, AQP-2, AQP-4-M1,AQP-4-M23, and AQP-5.

FIG. 3 depicts a Hummel-Dryer style assay for [3H]-labeled Compound 4(compound of formula 1a where R₁, R₃ and R₅ are each trifluoromethyl,and R₂, R₄ and R₆ are H) binding to purified AQP4b.

FIG. 4 depicts percent survival curves for the water toxicity mousemodel using 0.76 mg/kg Compound 1 (compound of formula 1a where R₁ ischloro, R₃ and R₅ are each trifluoromethyl, and R₂, R₄ and R₆ are H).

FIG. 5 depicts inhibition of cerebral edema formation in a mouse watertoxicity model determined by brain water content using Compound 1.

FIG. 6 depicts inhibition of cerebral edema formation by Compound 1 inthe mouse water toxicity model by MRI brain volume analysis, with n=14mice/treatment. A time course of edema formation is shown comparing nodrug vs. Compound 1 at 0.76 mg/kg. The first time point at 5.67 mincoincides with the scan slice at the middle of the brain during thefirst post-injection scan. Other time points are placed in a similarmanner. The data is fitted to a single exponential equation:V/V ₀ =V _(i) +dV _(max)(1−e ^((−kt)));where V/V₀=relative brain volume, V_(i)=initial relative brain volume,dV_(max)=maximum change in relative brain volume, k=first order rateconstant (min⁻¹), and t=time in minutes.

FIG. 7 depicts the calcein fluorescence end-point assay used for highthroughput screening

FIG. 8 depicts hit validation using the Cell Bursting Aquaporin Assay;inset shows the structure of Compound 3.

FIG. 9 depicts reduction in intracranial pressure (ICP) in the mousewater toxicity model with Compound 1 at 0.76 mg/kg.

FIG. 10 depicts plasma and serum levels of Compound 1 converted fromCompound 5 (compound of formula 1a where R₁ is chloro, R₃ and R₅ areeach trifluoromethyl, R₂ and R₄ are H, and R₆ is P(═O)(O)₂ in disodiumsalt form).

FIG. 11 depicts mouse middle cerebral artery occlusion (MCAo) model ofischemic stroke.

FIG. 12 depicts relative change in hemispheric brain volume in the mousemiddle cerebral artery occlusion (MCAo) model.

FIG. 13 depicts neurological outcome following MCAo in mice treated withsaline (no drug, •) or Compound 5 (o).

DETAILED DESCRIPTION OF THE INVENTION

The following description of the preferred embodiments is merelyexemplary in nature and is in no way intended to limit the invention,its application, or uses.

Expression of Aquaporin-4 (AQP4) is upregulated in animal models oftrauma, stroke and water intoxication as well as around human malignantbrain tumors. Aquaporin-4 (AQP4) has been shown to play a critical rolein the development of cerebral and spinal cord edema. AQP4 provides theprimary route for water movement across the BBB and glia limitans. AQP4knockout mice, without the APQ4 gene, have improved survival compared towild-type mice in models of ischemic stroke, water toxicity, bacterialmeningitis, and spinal cord compression.

Cerebral edema (CE) is generally divided into 2 major categories:vasogenic and cytotoxic. Vasogenic cerebral edema may occur when abreach in the blood-brain barrier (BBB) allows water and solutes todiffuse into the brain. It has been reported that AQP4-null mice haveincreased brain edema in a model of subarachnoid hemorrhage, suggestingthat AQP4 may be required for the clearance of water collected inintercellular space. In contrast, cytotoxic cerebral edema may beinitiated by ischemia which results in reduced plasma osmolality ratherthan a disrupted BBB. Ischemia may to a drop in ATP levels which isthought to slow the Na—K ATPase pump resulting in an uptake of Na⁺ andCl⁻ through leakage pathways. The net effect may be a cellular osmoticimbalance, drawing H₂O into cells—astrocytes more so than neurons—andleading to increased ICP. Mouse models for ischemic stroke, watertoxicity, bacterial meningitis, and spinal-cord compression fall intothis category. In these models, AQP4-null mice have reduced CE pointingto AQP4 as the central pathway for water movement into the brain duringthe formation of cytotoxic CE. However, cytotoxic and vasogenic edemaare not sharply divided categories; an injury that initially causescytotoxic edema may be followed later, e.g., within the next hours todays, by vasogenic edema. This may suggest different treatments forcerebral edema at different times.

It has been reported that propensity to epileptic seizures is related torelative cellular and extracellular space (ECS) volume.Hyperexcitability and increased epileptiform activity results fromhypotonic exposure which decreases ECS volume, while attenuatedepileptiform activity results from hyperosmolar medium. Furosemide,which blocks seizure-induced cell swelling, has been reported to inhibitepileptiform activity in vitro and in vivo. AQP4 knockout mice werereported to have lower seizure susceptibility to the convulsantpentylenetetrazol and a greater electrographic seizure threshold whenseizures were induced by electrical stimulation in the hippocampus. Itwas also reported that AQP4 knockout mice had more prolongedhippocampal-stimulation evoked seizures compared to wild type mice.

AQP4 is expressed in the Müller cells in the retina. Studies haveimplicated Müller cells in the pathogenesis of retinal injury afterischemia. It has been reported that AQP4 deletion in mice conferredsignificant preservation of retinal function and architecture afterretinal ischemia.

AQP4 is reportedly found in mammalian hearts. It has been reported thatAQP4 expression in the human heart is present at both the mRNA andprotein level. Water accumulates in the myocardium as a result ofischemia, when ischemic tissue becomes hyperosmolar and attracts waterfrom the capillary lumen. The water is transported into the myocardialcells, for example, into cardiomyocytes. Reperfusion deliversnormoosmolar blood to the hyperosmolar cells, which leads to furthercell swelling, which may even involve cells outside the risk area. Thiswater accumulation leads to a pronounced depression of cardiac function,and aggravates effects of shortage in oxygen and nutrient supply.Myocardial ischemia/reperfusion injury refers to damage caused byischemia followed by reperfusion in the heart. It has been reported thatAQP4 knockout mice had reduced infarct size after both ex vivoischemia-reperfusion and after in vivo ischemia without reperfusion. Itwas concluded that the AQP4 knockout genotype conferred increasedtolerance to ischemic injury.

Neuromyelitis optica (NMO) is a neuroinflammatory demyelinating diseasethat primarily affects optic nerve and spinal cord. A feature of NMO isthe presence of serum antibodies directed against extracellular epitopeson AQP4. It has been reported that most, if not all, NMO patients areseropositive for AQP4 autoantibodies (NMO-IgG). It is thought thatNMO-IgG binding to AQP4 in astrocytes initiates an inflammatory cascadeand the consequent neuroinflammation and myelin loss produceneurological deficits. Blocking binding of those antibodies to AQP4could prevent the initiation of the inflammatory cascade.

In one embodiment, the invention provides methods of treating edemamediated by aquaporin, e.g., AQP4, wherein the edema is consequent tohypoxia, e.g., general systemic hypoxia, e.g., hypoxia caused by aninterruption of blood perfusion, for example wherein the edema iscerebral edema consequent to hypoxia caused by cardiac arrest, stroke,or other interruption of blood perfusion to the brain, or wherein theedema is cardiac edema consequent to cardiac ischemia or otherinterruption of blood flow to the heart. Hypoxia can lead to developmentof metabolic acidosis (e.g. lactic acidosis), which in turn leads toedema, and the edema itself can then reduce blood perfusion, leading tocell death and poorer outcomes, particularly in tissues where swellingis physically constrained, for example within the skull or within thepericardium. This hypoxia is believed to be why, for example, patientswho have been rescued from cardiac arrest may subsequently exhibit brainswelling, as well as damage to the cardiac tissue. Blocking aquaporinchannels, e.g., AQP4, e.g., by administering an aquaporin-inhibitingcompound as described herein, inhibits or controls this edema, therebylimiting further damage to the affected tissue.

Aquaporin-2 (AQP2) is the primary route of water movement at thecollecting duct in the kidney. Blocking this water channel would lowerwater reabsorption without incurring electrolyte imbalances orinterfering with vasopressin receptor-mediated signaling. Evidence thatan AQP2 blocker would not produce electrolyte imbalances, and instead bean effective treatment for hyponatremia, comes from patients withdiabetes insipidus who lack functional AQP2. They exhibit chronicaquaresis but—if normal hydration is maintained—do not demonstrate anyother consequence of their long term loss of AQP2 function.

The invention thus provides the use of aquaporin inhibitors incontrolling diseases or conditions of water imbalance, including edema,particularly edema of the brain and spinal cord, e.g., following traumaor ischemic stroke, as well as the edema associated with glioma,meningitis, acute mountain sickness, epileptic seizures, infections,metabolic disorders, hypoxia, water intoxication, hepatic failure,hepatic encephalopathy, hypoxia, and diabetic ketoacidosis by inhibitingwater uptake through the BBB, and also useful in treating andcontrolling hyponatremia and excessive fluid retention, by inhibitingwater uptake at the kidneys. This invention also provides the use ofaquaporin inhibitors in controlling diseases or conditions includingepilepsy, retinal ischemia and other diseases of the eye associated withabnormalities in intraocular pressure and/or tissue hydration,myocardial ischemia, myocardial ischemia/reperfusion injury, myocardialinfarction, myocardial hypoxia, congestive heart failure, sepsis,neuromyelitis optica, and migraines.

In one embodiment, the invention provides a method (Method 1) oftreating or controlling a disease or condition mediated by an aquaporincomprising administering to a patient in need thereof an effectiveamount of a phenylbenzamide compound, e.g., an effective amount ofniclosamide (5-chloro-N-(2-chloro-4-nitrophenyl)-2-hydroxybenzamide) ora compound of Formula I:

wherein X represents a connecting group whose number of atoms in themain chain is 2 to 5 (said connecting group may be substituted), Arepresents a hydrogen atom or an acyl group which may be substituted, ora C₁ to C₆ alkyl group which may be substituted, or A may bind toconnecting group X to form a cyclic structure which may be substituted,E represents an aryl group which may be substituted or a heteroarylgroup which may be substituted, ring Z represents an arene which mayhave one or more substituents in addition to the group represented byformula —O-A wherein A has the same meaning as that defined above andthe group represented by formula —X-E wherein each of X and E has thesame meaning as that defined above, or a heteroarene which may have oneor more substituents in addition to the group represented by formula—O-A wherein A has the same meaning as that defined above and the grouprepresented by formula —X-E wherein each of X and E has the same meaningas that defined above; in free or pharmaceutically acceptable salt form,including solvate or hydrate forms; e.g.

-   1.1. Method 1 wherein the compound of Formula I is selected from the    compounds of formula (I) as described in US 2010/0274051 A1 or U.S.    Pat. No. 7,700,655, e.g., a compound of general formulas (I), (I-1),    (I-2), (I-3), and (I-4) as set forth therein, e.g., selected from    Compound Nos. 1-223 as set forth in US 2010/0274051 or Compound Nos.    301-555 as set forth in U.S. Pat. No. 7,700,655.-   1.2. Method 1.1 wherein the compound of Formula I is selected from    the free or pharmaceutically acceptable salt forms of:-   N-[3,5-bis(trifluoromethyl)phenyl]-5-fluoro-2-hydroxybenzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-5-cyano-2-hydroxybenzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-(trifluoromethyl)benzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-(1,1,2,2,2-pentafluoroethyl)benzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-5-(2,2-di    cyanoethen-1-yl)-2-hydroxybenzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-5-ethynyl-2-hydroxybenzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-(phenylethynyl)benzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-[(trimethylsilyl)ethynyl]benzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-4-hydroxybiphenyl-3-carboxamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-(3-thienyl)benzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-(1-pyrrolyl)benzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-(2-methylthiazol-4-yl)benzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-(2-pyridyl)benzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-5-dimethylsulfamoyl-2-hydroxybenzamide,-   N-[3,5-bis(trifluoromethyl)phenyl]-2-hydroxy-5-(pyrrole-1-sulfonyl)benzamide,-   N-[2,5-bis(trifluoromethyl)phenyl]-5-chloro-2-hydroxybenzamide,-   N-(2,5-bis(trifluoromethyl)phenyl-5-bromo-2-hydroxybenzamide,-   2-acetoxy-N-[2,5-bis(trifluoromethyl)phenyl]-5-chlorobenzamide,-   2-acetoxy-N-[3,5-bis(trifluoromethyl)phenyl]-5-chlorobenzamide,-   5-chloro-N-[2-fluoro-3-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-chloro-N-[2-fluoro-5-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-chloro-N-[2-chloro-5-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-bromo-N-[2-chloro-5-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   2-acetoxy-5-chloro-N-[2-chloro-5-(trifluoromethyl)phenyl]benzamide,-   5-chloro-N-[3-fluoro-5-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-bromo-N-[3-bromo-5-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-chloro-N-[3-fluoro-5-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-chloro-N-[4-fluoro-3-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-bromo-N-[4-chloro-3-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-chloro-2-hydroxy-N-[2-nitro-5-(trifluoromethyl)phenyl]benzamide,-   5-bromo-N-[4-cyano-3-(trifluoromethyl)phenyl]-2-hydroxybenzamide,-   5-chloro-2-hydroxy-N-[2-methyl-3-(trifluoromethyl)phenyl]benzamide,-   5-chloro-2-hydroxy-N-[2-methyl-5-(trifluoromethyl)phenyl]benzamide,-   2-hydroxy-5-methyl-N-[2-methyl-5-(trifluoromethyl)phenyl]benzamide,-   5-chloro-2-hydroxy-N-[4-methyl-3-(trifluoromethyl)phenyl]benzamide,-   2-hydroxy-5-methyl-N-[4-methyl-3-(trifluoromethyl)phenyl]benzamide,-   5-bromo-2-hydroxy-N-[2-methoxy-5-(trifluoromethyl)phenyl]benzamide,-   5-chloro-2-hydroxy-N-[2-methoxy-5-trifluoromethyl)phenyl]benzamide,-   5-bromo-2-hydroxy-N-[3-methoxy-5-(trifluoromethyl)phenyl]benzamide,-   5-chloro-2-hydroxy-N-[4-methoxy-3-(trifluoromethyl)phenyl]benzamide,-   5-chloro-2-hydroxy-N-[2-methylsulfanyl-5-(trifluoromethyl)phenyl]benzamide,-   5-chloro-2-hydroxy-N-[2-(1-pyrrolidino)-5-(trifluoromethyl)phenyl]benzamide,-   5-chloro-2-hydroxy-N-[2″-morpholino-5-(trifluoromethyl)phenyl]benzamide,-   5-bromo-N-[5-bromo-4-(trifluoromethyl)thiazol-2-yl]-2-hydroxybenzamide,-   5-chloro-N-{5-cyano-4-[(1,1-dimethyl)ethyl]thiazol-2-yl}-2-hydroxybenzamide,-   5-bromo-N-{5-cyano-4-[(1,1-dimethyl)ethyl]thiazol-2-yl}-2-hydroxybenzamide,-   2-(5-bromo-2-hydroxybenzoyl)amino-4    (trifluoromethyl)thiazol-5-carboxylic acid ethyl ester.-   1.3. Method 1 wherein A is C₁₋₄ acyl (e.g. acetyl).-   1.4. Method 1 wherein the compound is a compound of formula I as    described in U.S. Pat. No. 7,626,042, for example of formula I-1,    e.g. any of Compound Nos. 1-151 as described in U.S. Pat. No.    7,626,042.-   1.5. Method 1.4 wherein A is C₁₋₄ acyl (e.g. acetyl).-   1.6. Method 1 wherein A is the residue of an amino acid.-   1.7. Method 1 wherein A is a 5 to 6-membered non-aromatic    heterocyclic ring-carbonyl group, for example a 5 to 6-membered    non-aromatic heterocyclic ring-carbonyl group which comprises at    least one nitrogen atom as ring-constituting atoms (ring forming    atoms) of said heterocyclic ring and binds to the carbonyl group at    the nitrogen atom, e.g., wherein said 5 to 6-membered non-aromatic    heterocyclic ring is selected from 1-pyrrolidinyl group, piperidino    group, morpholino group, and 1-piperazinyl group, and said    heterocyclic ring may be substituted with one or more substituents,    e.g., independently selected from an alkyl group, an    alkyl-oxy-carbonyl group, and a carboxy group; for example wherein A    is (morpholin-4-yl)carbonyl.-   1.8. Method 1 wherein A is a N,N-di-substituted carbamoyl group,    wherein two substituents of said carbamoyl group may combine to each    other, together with the nitrogen atom to which they bind, to form a    nitrogen-containing heterocyclic group which may be substituted.-   1.9. Method 1 wherein A is (morpholin-4-yl)carbonyl.-   1.10. Method 1 wherein A is a phosphono group, which may be    substituted, e.g., dibenzyl phosphono, or unsubstituted.-   1.11. Method 1 wherein the compound of Formula I is a compound of    formula 1a:

-   -   wherein R₁, R₂, R₃, R₄, and R₅ are selected from H, halo,        halogenated C₁₋₄ alkyl (e.g., trifluoromethyl), and cyano; and        R₆ is selected from H and physiologically hydrolysable and        acceptable acyl, e.g., wherein R₆ is A as hereinbefore defined        in any of Methods 1-1.9;    -   in free, pharmaceutically acceptable salt form.

-   1.12. Method 1.11 wherein the compound of Formula I is a compound of    formula 1a wherein R₁ is selected from trifluoromethyl, chloro,    fluoro, and bromo; R₃ and R₅ are the same or different and selected    from trifluoromethyl, chloro, fluoro, and bromo; and R₂ and R₄ are    both H.

-   1.13. Method 1.12 wherein the compound of Formula I is a compound of    formula 1a wherein R₁ is selected from chloro and bromo; R₃ and R₅    are both trifluoromethyl; and R₂, R₄ and R₆ are all H, e.g., wherein    the compound of formula 1a is selected from:

-   1.14. Method 1.11 or 1.12 wherein R₆ is H.-   1.15. Method 1.11 or 1.12 wherein R₆ is acetyl.-   1.16. Method 1.11 or 1.12 wherein the compound of Formula I is a    compound of formula 1a wherein R₁ is selected from chloro and bromo;    R₃ and R₅ are both trifluoromethyl; and R₂ and R₄ are H and R₆ is    acetyl, e.g., wherein the compound of formula 1a is selected from:

-   1.17. Method 1.13 wherein the compound of formula 1a is Compound 1

-   1.18. Method 1.12 wherein the compound of Formula I is a compound of    formula 1a wherein R₁, R₃ and R₅ are each chloro, and R₂, R₄ and R₆    are each H (Compound 3).-   1.19. Method 1.12 wherein the compound of Formula I is a compound of    formula 1a wherein R₁, R₃ and R₅ are each trifluoromethyl, and R₂,    R₄ and R₆ are each H (Compound 4).-   1.20. Method 1.11 or 1.12 wherein the compound of Formula I is a    compound of formula 1a, and R₆ is C₁₋₄ acyl (e.g. acetyl).-   1.21. Method 1.11 or 1.12 wherein the compound of Formula I is a    compound of formula 1a, and R₆ is the residue of an amino acid.-   1.22. Method 1.11 or 1.12 wherein the compound of Formula I is a    compound of formula 1a, and R₆ is a 5 to 6-membered non-aromatic    heterocyclic ring-carbonyl group, for example a 5 to 6-membered    non-aromatic heterocyclic ring-carbonyl group which comprises at    least one nitrogen atom as ring-constituting atoms (ring forming    atoms) of said heterocyclic ring and binds to the carbonyl group at    the nitrogen atom, e.g., wherein said 5 to 6-membered non-aromatic    heterocyclic ring is selected from 1-pyrrolidinyl group, piperidino    group, morpholino group, and 1-piperazinyl group, and said    heterocyclic ring may be substituted with one or more substituents,    e.g., independently selected from an alkyl group, an    alkyl-oxy-carbonyl group, and a carboxy group; for example wherein    R₆ is (morpholin-4-yl)carbonyl.-   1.23. Method 1.11 or 1.12 wherein the compound of Formula I is a    compound of formula 1a, and R₆ is a N,N-di-substituted carbamoyl    group, wherein two substituents of said carbamoyl group may combine    to each other, together with the nitrogen atom to which they bind,    to form a nitrogen-containing heterocyclic group which may be    substituted.-   1.24. Method 1.11 or 1.12 wherein the compound of Formula I is a    compound of formula 1a, and R₆ is a (morpholin-4-yl)carbonyl group.-   1.25. Method 1.11 or 1.12 wherein the compound of Formula I is a    compound of formula 1a, and R₆ is a phosphono group, which may be    substituted, e.g. dibenzylphosphono, or unsubstituted.-   1.26. Method 1.25 wherein the compound of formula 1a is selected    from:

-   1.27. Method 1.26 wherein the compound of formula 1a is Compound 5

-   1.28. Method 1 wherein the phenylbenzamide compound is niclosamide    or the compound shown below

-   1.29. Any of Method 1, et seq. wherein the aquaporin is AQP4.-   1.30. Any of Method 1, et seq. wherein the condition to be treated    or controlled is selected from edema, e.g. edema of the brain or    spinal cord, e.g., cerebral edema, e.g. cerebral edema consequent to    head trauma, ischemic stroke, glioma, meningitis, acute mountain    sickness, epileptic seizures, infections, metabolic disorders, water    intoxication, hepatic failure, hepatic encephalopathy, or diabetic    ketoacidosis or, e.g., spinal cord edema, e.g., spinal cord edema    consequent to spinal cord trauma, e.g., spinal cord compression.-   1.31. Method 1.30 further comprising a treatment selected from one    or more of the following: optimal head and neck positioning to    facilitate venous outflow, e.g. head elevation 30°; avoidance of    dehydration; systemic hypotension; maintenance of normothermia or    hypothermia; aggressive measures; osmotherapy, e.g., using mannitol    or hypertonic saline; hyperventilation; therapeutic pressor therapy    to enhance cerebral perfusion; administration of barbiturates to    reduce cerebral metabolism (CMO₂); hemicraniectomy; administration    of aspirin; administration of amantadine; intravenous thrombolysis    (e.g. using rtPA); mechanical clot removal; angioplasty; and/or    stents.-   1.32. Any of Method 1, et seq. wherein the patient is at elevated    risk of cerebral edema, e.g., due to head trauma, ischemic stroke,    glioma, meningitis, acute mountain sickness, epileptic seizure,    infection, metabolic disorder, water intoxication, hepatic failure,    hepatic encephalopathy, or diabetic ketoacidosis.-   1.33. Method 1.30 wherein the patient has suffered a stroke, head    injury, or spinal injury.-   1.34. Method 1.33 wherein the patient has suffered a stroke, head    injury or spinal injury within 12 hours, e.g. within 6 hours,    preferably within 3 hours of commencing treatment.-   1.35. Method 1.30 wherein the patient is at elevated risk of    suffering a stroke, head injury or spinal injury, e.g., in combat or    in an athletic competition.-   1.36. Any of Method 1, et seq. wherein the patient already has    cerebral edema.-   1.37. Any of Method 1, et seq. wherein the condition to be treated    or controlled is cerebral edema consequent to a stroke or a    traumatic brain injury.-   1.38. Any of Method 1, et seq. wherein the condition to be treated    or controlled is cerebral edema consequent to a middle cerebral    artery stroke.-   1.39. Any of Method 1, et seq. wherein the condition to be treated    or controlled is cerebral edema consequent to closed head trauma.-   1.40. Any of Methods 1-1.32 wherein the condition to be treated or    controlled is cerebral edema consequent to an epileptic seizure.-   1.41. Any of Methods 1-1.32 wherein the condition to be treated or    controlled is cerebral edema consequent to an infection.-   1.42. Any of Methods 1-1.32 wherein the condition to be treated or    controlled is cerebral edema consequent to a metabolic disorder.-   1.43. Any of Methods 1-1.32 wherein the condition to be treated or    controlled is cerebral edema consequent to glioma.-   1.44. Any of Methods 1-1.32 wherein the condition to be treated or    controlled is cerebral edema consequent to meningitis, acute    mountain sickness, or water intoxication.-   1.45. Any of Methods 1-1.32 wherein the condition to be treated or    controlled is cerebral edema consequent to hepatic failure, hepatic    encephalopathy, or diabetic ketoacidosis.-   1.46. Any of Methods 1-1.31 wherein the condition to be treated or    controlled is cerebral edema consequent to an abscess.-   1.47. Any of Methods 1-1.31 wherein the condition to be treated or    controlled is cerebral edema consequent to eclampsia.-   1.48. Any of Methods 1-1.31 wherein the condition to be treated or    controlled is cerebral edema consequent to Creutzfeldt-Jakob    disease.-   1.49. Any of Methods 1-1.31 wherein the condition to be treated or    controlled is cerebral edema consequent to lupus cerebritis.-   1.50. Any of Methods 1-1.31 wherein the condition to be treated or    controlled is edema consequent to hypoxia, e.g., general systemic    hypoxia, e.g., hypoxia caused by an interruption of blood perfusion,    for example wherein the edema is cerebral edema consequent to    hypoxia caused by cardiac arrest, stroke, or other interruption of    blood perfusion to the brain, or wherein the edema is cardiac edema    consequent to cardiac ischemia or other interruption of blood flow    to the heart.-   1.51. Any of Methods 1-1.31 wherein the condition to be treated or    controlled is cerebral and/or optic nerve edema consequent to    microgravity and/or radiation exposure, e.g., exposure from space    flight or from working with radioactive materials or from working in    radioactive areas.-   1.52. Any of Methods 1-1.31 wherein the condition to be treated or    controlled is cerebral edema consequent to an invasive central    nervous system procedures, e.g., neurosurgery, endovascular clot    removal, spinal tap, aneurysm repair, or deep brain stimulation.-   1.53. Method 1.51 or 1.52 wherein the patient is at elevated risk of    edema, e.g., due to microgravity and/or radiation exposure,    neurosurgery, endovascular clot removal, spinal tap, aneurysm    repair, or deep brain stimulation.-   1.54. Method 1.51 or 1.52 wherein the patient already has edema.-   1.55. Any of Methods 1, et seq. wherein the edema is cytotoxic    cerebral edema or is primarily cytotoxic cerebral edema.-   1.56. Any of Methods 1-1.45 or 1.50 wherein the edema is cytotoxic    cerebral edema or is primarily cytotoxic cerebral edema.-   1.57. Any of Methods 1-1.30 wherein the condition to be treated or    controlled is spinal cord edema, e.g., spinal cord edema consequent    to a spinal cord trauma, e.g., spinal cord compression.-   1.58. Method 1.57 wherein the condition to be treated or controlled    is spinal cord edema consequent to spinal cord compression.-   1.59. Any of Methods 1-1.30 wherein the condition to be treated or    controlled is retinal edema.-   1.60. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is epilepsy.-   1.61. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is retinal ischemia or other diseases of the eye    associated with abnormalities in intraocular pressure and/or tissue    hydration.-   1.62. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is myocardial ischemia.-   1.63. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is myocardial ischemia/reperfusion injury.-   1.64. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is myocardial infarction.-   1.65. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is myocardial hypoxia.-   1.66. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is congestive heart failure.-   1.67. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is sepsis.-   1.68. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is a migraine.-   1.69. Any of Methods 1-1.28 wherein the aquaporin is AQP2.-   1.70. Any of Methods 1-1.28 or 1.69 wherein the condition to be    treated is hyponatremia or excessive fluid retention, e.g.,    consequent to heart failure (HF), for example congestive heart    failure, liver cirrhosis, nephrotic disorder, or syndrome of    inappropriate antidiuretic hormone secretion (SIADH).-   1.71. Any of Methods 1-1.28 or 1.69-1.70 further comprising one or    more of restriction of dietary sodium, fluid and/or alcohol; and/or    administration of one or more diuretics, vasopressin receptor    antagonists, angiotensin converting enzyme (ACE) inhibitors,    aldosterone inhibitors, angiotensin receptor blockers (ARBs),    beta-adrenergic antagonists (beta-blockers), and/or digoxin.-   1.72. Any of Method 1, et seq. wherein the niclosamide or the    compound of Formula I or formula 1a inhibits aquaporin activity,    e.g., AQP2 and/or AQP4 activity, by at least 50% at concentrations    of 10 micromolar or less, for example inhibits APQ2 and/or AQP4    activity by at least 50% at concentrations of 10 micromolar or less    in an aquaporin-mediated cell volume change assay, e.g., is active    in any of the assays of Method 10, et seq. infra.-   1.73. Any of Methods 1-1.29 wherein the condition to be treated or    controlled is neuromyelitis optica.-   1.74. Any of Method 1, et seq. wherein the niclosamide or the    compound of Formula I or formula 1a is administered orally.-   1.75. Any of Method 1, et seq. wherein the niclosamide or the    compound of Formula I or formula 1a is administered parenterally.-   1.76. Method 1.75 wherein the niclosamide or the compound of Formula    I or formula 1a is administered intravenously.-   1.77. Any of Method 1, et seq. wherein the patient is human.-   1.78. Any of Method 1, et seq. wherein the onset of action of any of    the compounds identified in Methods 1-1.28 is fairly rapid.

The invention further provides a phenylbenzamide, e.g. niclosamide or acompound of Formula I or formula 1a as hereinbefore described, for usein treating or controlling a disease or condition mediated by anaquaporin, e.g., in any of Methods 1, 1.1, et seq.

The invention further provides a phenylbenzamide, e.g. niclosamide or acompound of Formula I or formula 1a as hereinbefore described, in themanufacture of a medicament for treating or controlling a disease orcondition mediated by an aquaporin, e.g., for use in any of Methods 1,1.1, et seq.

The invention further provides a pharmaceutical composition comprising aphenylbenzamide, e.g. niclosamide or a compound of Formula I or formula1a as hereinbefore described, in combination with a pharmaceuticallyacceptable diluent or carrier for use in treating or controlling adisease or condition mediated by an aquaporin, e.g., in any of Methods1, 1.1, et seq.

Phenylbenzamides, e.g. of Formula I or formula 1a as hereinbeforedescribed, may exist in free or salt form, e.g., as acid addition salts.In this specification unless otherwise indicated language such as“compound of Formula I or formula 1a” or “compounds of Formula I orformula 1a” is to be understood as embracing the compounds in any form,for example free base or acid addition salt form. Pharmaceuticallyacceptable salts are known in the art and include salts which arephysiologically acceptable at the dosage amount and form to beadministered, for example hydrochlorides.

Examples of the acyl group include, for example, formyl, glyoxyloylgroup, thioformyl group, carbamoyl group, thiocarbamoyl group, sulfamoylgroup, sulfinamoyl group, carboxy group, sulfo group, phosphono group,and groups represented by the following formulas:

wherein R^(a1) and R^(b1) may be the same or different and represent ahydrocarbon group or a heterocyclic group, or R^(a1) and R^(b1) combineto each other, together with the nitrogen atom to which they bind, toform a cyclic amino group. Acyl includes physiologically hydrolysableand acceptable acyl group. Examples of the acyl group of “an acyl groupwhich may be substituted” as used herein, e.g., in relation to “A”include similar groups to the acyl group in the aforementioneddefinition. “A” is a group selected from the following substituent groupω:[Substitutent group ω] a hydrocarbon-carbonyl group which may besubstituted, a heterocyclic ring-carbonyl group which may besubstituted, a hydrocarbon-oxy-carbonyl group which may be substituted,a hydrocarbon-sulfonyl group which may be substituted, a sulfamoyl groupwhich may be substituted, a sulfo group which may be substituted, aphosphono group which may be substituted, and a carbamoyl group whichmay be substituted. By the term “physiologically hydrolysable andacceptable acyl” as used herein, e.g., in relation to “A” or “R₆” incompounds of Formula I or formula 1a, is meant a residue of an acid, forexample a carboxylic acid, a carbamic acid or a phosphoric acid (e.g.,optionally substituted carbonyl such as acetyl or the residue of anamino acid, optionally substituted carbamoyl, e.g.(morpholin-4-yl)carbonyl, or optionally substituted phosphono e.g.,dibenzylphosphono), linked to an oxygen, e.g., as depicted in Formula 1or formula 1a above, e.g. to form an ester or phosphoester with acompound of Formula I or formula 1a, which is capable of hydrolysis fromsaid oxygen under physiological conditions to yield an acid which isphysiologically tolerable at doses to be administered, together with thecorresponding hydroxy compound of Formula I or formula 1a wherein A orR₆ is H. As will be appreciated the term thus embraces conventionalpharmaceutical prodrug forms, although it is not necessarily requiredthat the compounds must be hydrolyzed in order to be active. The acylcompounds may be prepared by conventional means, e.g., by acylation of acompound of Formula 1 or formula 1a, wherein A or R₆ is H, with thedesired acid or acid halide. Examples of acylated compounds and methodsof making them are provided, e.g., in US 2010/0274051 A1, U.S. Pat. No.7,700,655, and in U.S. Pat. No. 7,626,042, each incorporated herein byreference.

The term “patient” includes human or non-human (i.e., animal) patient.In a particular embodiment, the invention encompasses both human andnonhuman. In another embodiment, the invention encompasses nonhuman. Inanother embodiment, the term encompasses human.

The term “fairly rapid” with respect to onset of action means that thetime it takes after a compound is administered for a response to beobserved is 30 minutes or less, for example 20 minutes or less, forexample or 15 minutes or less, for example 10 minutes or less, forexample 5 minutes or less, for example 1 minute or less.

Phenylbenzamides, e.g. of Formula 1 or formula 1a as hereinbeforedescribed for use in the methods of the invention may be used as a soletherapeutic agent, but may also be used in combination or forco-administration with other active agents, for example in conjunctionwith conventional therapies for cerebral edema, stroke, traumatic braininjury, glioma, meningitis, acute mountain sickness, infection,metabolic disorder, hypoxia, water intoxication, hepatic failure,hepatic encephalopathy, diabetic ketoacidosis, abscess, eclampsia,Creutzfeldt-Jakob disease, lupus cerebritis, edema of the optic nerve,hyponatremia, fluid retention, epilepsy, retinal ischemia or otherdiseases of the eye associated with abnormalities in intraocularpressure and/or tissue hydration, myocardial ischemia, myocardialischemia/reperfusion injury, myocardial infarction, myocardial hypoxia,congestive heart failure, sepsis, neuromyelitis optica, or migraines.

In a further embodiment, the invention provides a method (Method 2) oftreating or controlling edema, e.g. edema of the brain or spinal cord,e.g., cerebral edema, e.g. cerebral edema consequent to head trauma,ischemic stroke, glioma, meningitis, acute mountain sickness, epilepticseizures, infections, metabolic disorders, hypoxia, water intoxication,hepatic failure, hepatic encephalopathy, diabetic ketoacidosis, abscess,eclampsia, Creutzfeldt-Jakob disease, or lupus cerebritis, as well asedema consequent to microgravity and/or radiation exposure, as well asedema consequent to invasive central nervous system procedures, e.g.,neurosurgery, endovascular clot removal, spinal tap, aneurysm repair, ordeep brain stimulation or, e.g., retinal edema or, e.g., spinal cordedema, e.g., spinal cord edema consequent to spinal cord trauma, e.g.,spinal cord compression, comprising administering an effective amount ofan inhibitor of AQP4, e.g, a compound binding to AQP4, to a patient inneed thereof, e.g., wherein the inhibitor of AQP4 is selected fromphenylbenzamides, e.g. niclosamide or a compound of Formula I or formula1a as hereinbefore described, e.g., any of the compounds identified inMethods 1-1.28 above, for example

2.

-   -   2.1. Method 2 further comprising a treatment selected from one        or more of the following: optimal head and neck positioning to        facilitate venous outflow, e.g. head elevation 30°; avoidance of        dehydration; systemic hypotension; maintenance of normothermia        or hypothermia; aggressive measures; osmotherapy, e.g., using        mannitol or hypertonic saline; hyperventilation; therapeutic        pressor therapy to enhance cerebral perfusion; administration of        barbiturates to reduce of cerebral metabolism (CMO₂);        hemicraniectomy; administration of aspirin; administration of        amantadine; intravenous thrombolysis (e.g. using rtPA);        mechanical clot removal; angioplasty; and/or stents.    -   2.2. Method 2 or 2.1 wherein the patient is at elevated risk of        cerebral edema, e.g., due to head trauma, ischemic stroke,        glioma, meningitis, acute mountain sickness epileptic seizure,        infection, metabolic disorder, water intoxication, hepatic        failure, hepatic encephalopathy, or diabetic ketoacidosis.    -   2.3. Method 2, 2.1, or 2.2 wherein the patient has suffered a        stroke, head injury, or spinal injury.    -   2.4. Any of Method 2, et seq. wherein the patient has suffered a        stroke, head injury or spinal injury within 12 hours, e.g.        within 6 hours, preferably within 3 hours of commencing        treatment.    -   2.5. Any of Method 2, et seq. wherein the patient is at elevated        risk of suffering a stroke, head injury or spinal injury, e.g.,        in combat or in an athletic competition.    -   2.6. Any of Method 2, et seq. wherein the patient already has        cerebral edema.    -   2.7. Any of Method 2, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a stroke        or a traumatic brain injury.    -   2.8. Any of Method 2, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a middle        cerebral artery stroke.    -   2.9. Any of Method 2, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a closed        head trauma.    -   2.10. Any of Methods 2-2.2 wherein the condition to be treated        or controlled is cerebral edema consequent to an epileptic        seizure.    -   2.11. Any of Methods 2-2.2 wherein the condition to be treated        or controlled is cerebral edema consequent to an infection.    -   2.12. Any of Methods 2-2.2 wherein the condition to be treated        or controlled is cerebral edema consequent to a metabolic        disorder.    -   2.13. Any of Methods 2-2.2 wherein the condition to be treated        or controlled is cerebral edema consequent to glioma.    -   2.14. Any of Methods 2-2.2 wherein the condition to be treated        or controlled is cerebral edema consequent to meningitis, acute        mountain sickness, or water intoxication.    -   2.15. Any of Methods 2-2.2 wherein the condition to be treated        or controlled is cerebral edema consequent to hepatic failure,        hepatic encephalopathy, or diabetic ketoacidosis.    -   2.16. Method 2 or 2.1 wherein the condition to be treated or        controlled is cerebral edema consequent to an abscess.    -   2.17. Method 2 or 2.1 wherein the condition to be treated or        controlled is cerebral edema consequent to eclampsia.    -   2.18. Method 2 or 2.1 wherein the condition to be treated or        controlled is cerebral edema consequent to Creutzfeldt-Jakob        disease.    -   2.19. Method 2 or 2.1 wherein the condition to be treated or        controlled is cerebral edema consequent to lupus cerebritis.    -   2.20. Method 2 or 2.1 wherein the condition to be treated or        controlled is cerebral and/or optic nerve edema consequent to        microgravity exposure, e.g., exposure from space flight or from        working with radioactive materials or from working in        radioactive areas.    -   2.21. Method 2 or 2.1 wherein the condition to be treated or        controlled is cerebral edema consequent to invasive central        nervous system procedures, e.g., neurosurgery, endovascular clot        removal, spinal tap, aneurysm repair, or deep brain stimulation.    -   2.22. Method 2.20 or 2.21 wherein the patient is at elevated        risk of edema, e.g., due to microgravity and/or radiation        exposure, neurosurgery, endovascular clot removal, spinal tap,        aneurysm repair, or deep brain stimulation.    -   2.23. Method 2.20 or 2.21 wherein the patient already has edema.    -   2.24. Any of Methods 2, et seq. wherein the edema is cytotoxic        cerebral edema or is primarily cytotoxic cerebral edema.    -   2.25. Any of Methods 2-2.15, et seq. wherein the edema is        cytotoxic cerebral edema or is primarily cytotoxic cerebral        edema.    -   2.26. Method 2 wherein the condition to be treated or controlled        is spinal cord edema, e.g., spinal cord edema consequent to        spinal cord trauma, e.g., spinal cord compression.    -   2.27. Method 2.26 wherein the condition to be treated or        controlled is spinal cord edema consequent to spinal cord        compression.    -   2.28. Method 2 wherein the condition to be treated or controlled        is retinal edema.    -   2.29. Any of Method 2, et seq. wherein the AQP4 inhibitor        inhibits AQP4 activity by at least 50% at concentrations of 10        micromolar or less, for example inhibits AQP4 activity by at        least 50% at concentrations of 10 micromolar or less in an        aquaporin-mediated cell volume change assay, e.g., is active in        any of the assays of Method 10, et seq. infra    -   2.30. Any of Method 2, et seq. wherein the duration of treatment        with an AQP4 inhibitor is less than 21 days, e.g., less than 2        weeks, e.g., one week or less.    -   2.31. Any of Method 2, et seq. wherein the AQP4 inhibitor is        administered orally.    -   2.32. Any of Method 2, et seq. wherein the AQP4 inhibitor is        administered parenterally.    -   2.33. Method 2.32 wherein the AQP4 inhibitor is administered        intravenously.    -   2.34. Any of Method 2, et seq. wherein the patient is human.    -   2.35. Any of Method 2, et seq. wherein the onset of action of        any of the compounds identified in Methods 1-1.28 is fairly        rapid.    -   2.36. Any of Method 2, et seq. wherein the edema is consequent        to hypoxia, e.g., general systemic hypoxia, e.g., hypoxia caused        by an interruption of blood perfusion, for example wherein the        edema is cerebral edema consequent to hypoxia caused by cardiac        arrest or other interruption of blood perfusion to the brain.

In a further embodiment, the invention provides a method (Method 3) oftreating or controlling a condition selected from hyponatremia andexcessive fluid retention, e.g., consequent to heart failure (HF), forexample congestive heart failure, liver cirrhosis, nephrotic disorder,or syndrome of inappropriate antidiuretic hormone secretion (SIADH)comprising administering an effective amount of an inhibitor of AQP2,e.g., a compound binding to AQP2, e.g., to a patient in need thereof,e.g., wherein the inhibitor of AQP2 is selected from phenylbenzamides,e.g. niclosamide or a compound of Formula I or formula 1a ashereinbefore described, e.g., any of the compounds identified in Methods1-1.28 above, for example

3.

-   -   3.1. Method 3 further comprising one or more of restriction of        dietary sodium, fluid and/or alcohol; and/or administration of        one or more diuretics, vasopressin receptor antagonists,        angiotensin converting enzyme (ACE) inhibitors, aldosterone        inhibitors, angiotensin receptor blockers (ARBs),        beta-adrenergic antagonists (beta-blockers), and/or digoxin.    -   3.2. Any of Method 3, et seq. wherein the AQP2 inhibitor        inhibits AQP2 activity by at least 50% at concentrations of 10        micromolar or less, for example inhibits APQ2 activity by at        least 50% at concentrations of 10 micromolar or less in an        aquaporin-mediated cell volume change assay, e.g., is active in        any of the assays of Method 10, et seq. infra    -   3.3. Any of Method 3, et seq. wherein the AQP2 inhibitor is        administered orally.    -   3.4. Any of Method 3, et seq. wherein the AQP2 inhibitor is        administered parenterally.    -   3.5. Method 3.4 wherein the AQP2 inhibitor is administered        intravenously.    -   3.6. Any of Method 3, et seq. wherein the patient is human.    -   3.7. Any of Method 3, et seq. wherein the onset of action of any        of the compounds identified in Methods 1-1.28 is fairly rapid.

In a further embodiment, the invention provides a method (Method 4) oftreating or controlling a condition selected from epilepsy, retinalischemia or other diseases of the eye associated with abnormalities inintraocular pressure and/or tissue hydration, myocardial ischemia,myocardial ischemia/reperfusion injury, myocardial infarction,myocardial hypoxia, congestive heart failure, sepsis, neuromyelitisoptica, or migraines comprising administering an effective amount of aninhibitor of AQP4, e.g, a compound binding to AQP4, to a patient in needthereof, e.g., wherein the inhibitor of AQP4 is selected fromphenylbenzamides, e.g. niclosamide or a compound of Formula I or formula1a as hereinbefore described, e.g., any of the compounds identified inMethod 1-1.28 above, for example

4.

-   -   4.1. Method 4 wherein the condition to be treated or controlled        is retinal ischemia or other diseases of the eye associated with        abnormalities in intraocular pressure and/or tissue hydration.    -   4.2. Method 4 wherein the condition to be treated or controlled        is myocardial ischemia.    -   4.3. Method 4 wherein the condition to be treated or controlled        is myocardial ischemia/reperfusion injury.    -   4.4. Method 4 wherein the condition to be treated or controlled        is myocardial infarction.    -   4.5. Method 4 wherein the condition to be treated or controlled        is myocardial hypoxia.    -   4.6. Method 4 wherein the condition to be treated or controlled        is congestive heart failure.    -   4.7. Method 4 wherein the condition to be treated or controlled        is sepsis.    -   4.8. Method 4 wherein the condition to be treated or controlled        is neuromyelitis optica.    -   4.9. Method 4 wherein the condition to be treated or controlled        is a migraine.    -   4.10. Any of Method 4, et seq. wherein the AQP4 inhibitor        inhibits AQP4 activity by at least 50% at concentrations of 10        micromolar or less, for example inhibits APQ4 activity by at        least 50% at concentrations of 10 micromolar or less in an        aquaporin-mediated cell volume change assay, e.g., is active in        any of the assays of Method 10, et seq. infra    -   4.11. Any of Method 4, et seq. wherein the AQP4 inhibitor is        administered orally.    -   4.12. Any of Method 4, et seq. wherein the AQP4 inhibitor is        administered parenterally.    -   4.13. Method 4.12 wherein the AQP4 inhibitor is administered        intravenously.    -   4.14. Any of Method 4, et seq. wherein the patient is human.    -   4.15. Any of Method 4, et seq. wherein the onset of action of        any of the compounds identified in Methods 1-1.28 is fairly        rapid.

In a further embodiment, the invention provides a method (Method 5) oftreating or controlling a disease or condition mediated by an aquaporincomprising administering to a patient in need thereof a phenylbenzamide,e.g. niclosamide or a compound of Formula I or formula 1a ashereinbefore described, e.g., any of the compounds identified in Method1-1.28 above, in an amount effective to inhibit the aquaporin, forexample

5.

-   -   5.1. Method 5 wherein the aquaporin is AQP4.    -   5.2. Method 5 or 5.1 wherein the condition to be treated or        controlled is selected from edema, e.g. edema of the brain or        spinal cord, e.g., cerebral edema, e.g. cerebral edema        consequent to head trauma, ischemic stroke, glioma, meningitis,        acute mountain sickness, epileptic seizures, infections,        metabolic disorders, water intoxication, hepatic failure,        hepatic encephalopathy, or diabetic ketoacidosis or, e.g.,        spinal cord edema, e.g., spinal cord edema consequent to spinal        cord trauma, e.g., spinal cord compression.    -   5.3. Method 5.2 further comprising a treatment selected from one        or more of the following: optimal head and neck positioning to        facilitate venous outflow, e.g. head elevation 30°; avoidance of        dehydration; systemic hypotension; maintenance of normothermia        or hypothermia; aggressive measures; osmotherapy, e.g., using        mannitol or hypertonic saline; hyperventilation; therapeutic        pressor therapy to enhance cerebral perfusion; administration of        barbiturates to reduce of cerebral metabolism (CMO₂);        hemicraniectomy; administration of aspirin; administration of        amantadine; intravenous thrombolysis (e.g. using rtPA);        mechanical clot removal; angioplasty; and/or stents.    -   5.4. Any of Method 5, et seq. wherein the patient is at elevated        risk of cerebral edema, e.g., due to head trauma, ischemic        stroke, glioma, meningitis, acute mountain sickness, epileptic        seizure, infection, metabolic disorder, water intoxication,        hepatic failure, hepatic encephalopathy, or diabetic        ketoacidosis.    -   5.5. Any of Method 5, et seq. wherein the patient has suffered a        stroke, head injury, or spinal injury.    -   5.6. Any of Method 5, et seq. wherein the patient has suffered a        stroke, head injury or spinal injury within 12 hours, e.g.        within 6 hours, preferably within 3 hours of commencing        treatment.    -   5.7. Any of Method 5, et seq. wherein the patient is at elevated        risk of suffering a stroke, head injury or spinal injury, e.g.,        in combat or in an athletic competition.    -   5.8. Any of Method 5, et seq. wherein the patient already has        cerebral edema.    -   5.9. Any of Method 5, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a stroke        or a traumatic brain injury.    -   5.10. Any of Method 5, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a middle        cerebral artery stroke.    -   5.11. Any of Method 5, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a closed        head trauma.    -   5.12. Any of Methods 5-5.4 wherein the condition to be treated        or controlled is cerebral edema consequent to an epileptic        seizure.    -   5.13. Any of Methods 5-5.4 wherein the condition to be treated        or controlled is cerebral edema consequent to an infection.    -   5.14. Any of Methods 5-5.4 wherein the condition to be treated        or controlled is cerebral edema consequent to a metabolic        disorder.    -   5.15. Any of Methods 5-5.4 wherein the condition to be treated        or controlled is cerebral edema consequent to glioma.    -   5.16. Any of Methods 5-5.4 wherein the condition to be treated        or controlled is cerebral edema consequent to meningitis, acute        mountain sickness, or water intoxication.    -   5.17. Any of Methods 5-5.4 wherein the condition to be treated        or controlled is cerebral edema consequent to hepatic failure,        hepatic encephalopathy, or diabetic ketoacidosis.    -   5.18. Any of Methods 5-5.3 wherein the condition to be treated        or controlled is cerebral edema consequent to an abscess.    -   5.19. Any of Methods 5-5.3 wherein the condition to be treated        or controlled is cerebral edema consequent to eclampsia.    -   5.20. Any of Methods 5-5.3 wherein the condition to be treated        or controlled is cerebral edema consequent to Creutzfeldt-Jakob        disease.    -   5.21. Any of Methods 5-5.3 wherein the condition to be treated        or controlled is cerebral edema consequent to lupus cerebritis.    -   5.22. Any of Methods 5-5.3 wherein the condition to be treated        or controlled is edema consequent to hypoxia, e.g., general        systemic hypoxia, e.g., hypoxia caused by an interruption of        blood perfusion, for example wherein the edema is cerebral edema        consequent to hypoxia caused by cardiac arrest, stroke, or other        interruption of blood perfusion to the brain, or wherein the        edema is cardiac edema consequent to cardiac ischemia or other        interruption of blood flow to the heart.    -   5.23. Any of Methods 5-5.3 wherein the condition to be treated        or controlled is cerebral and/or optic nerve edema consequent to        microgravity and/or radiation exposure, e.g., exposure from        space flight or from working with radioactive materials or from        working in radioactive areas.    -   5.24. Any of Methods 5-5.3 wherein the condition to be treated        or controlled is cerebral edema consequent to invasive central        nervous system procedures, e.g., neurosurgery, endovascular clot        removal, spinal tap, aneurysm repair, or deep brain stimulation.    -   5.25. Method 5.23 or 5.24 wherein the patient is at elevated        risk of edema, e.g., due to microgravity and/or radiation        exposure, neurosurgery, endovascular clot removal, spinal tap,        aneurysm repair, or deep brain stimulation.    -   5.26. Method 5.23 or 5.24 wherein the patient already has edema.    -   5.27. Any of Methods 5, et seq. wherein the edema is cytotoxic        cerebral edema or is primarily cytotoxic cerebral edema.    -   5.28. Any of Methods 5-5.17 or 5.22 wherein the edema is        cytotoxic cerebral edema or is primarily cytotoxic cerebral        edema.    -   5.29. Method 5 or 5.1 wherein the condition to be treated or        controlled is spinal cord edema, e.g., spinal cord edema        consequent to spinal cord trauma, e.g., spinal cord compression.    -   5.30. Method 5.29 wherein the condition to be treated or        controlled is spinal cord edema consequent to spinal cord        compression.    -   5.31. Any of Methods 5-5.2 wherein the condition to be treated        or controlled is retinal edema.    -   5.32. Method 5 or 5.1 wherein the condition to be treated or        controlled is epilepsy.    -   5.33. Method 5 or 5.1 wherein the condition to be treated or        controlled is retinal ischemia or other diseases of the eye        associated with abnormalities in intraocular pressure and/or        tissue hydration.    -   5.34. Method 5 or 5.1 wherein the condition to be treated or        controlled is myocardial ischemia.    -   5.35. Method 5 or 5.1 wherein the condition to be treated or        controlled is myocardial ischemia/reperfusion injury.    -   5.36. Method 5 or 5.1 wherein the condition to be treated or        controlled is myocardial infarction.    -   5.37. Method 5 or 5.1 wherein the condition to be treated or        controlled is myocardial hypoxia.    -   5.38. Method 5 or 5.1 wherein the condition to be treated or        controlled is congestive heart failure.    -   5.39. Method 5 or 5.1 wherein the condition to be treated or        controlled is sepsis.    -   5.40. Method 5 or 5.1 wherein the condition to be treated or        controlled is a migraine.    -   5.41. Method 5 wherein the aquaporin is AQP2.    -   5.42. Method 5 or 5.41 wherein the condition to be treated is        hyponatremia or excessive fluid retention, e.g., consequent to        heart failure (HF), for example congestive heart failure, liver        cirrhosis, nephrotic disorder, or syndrome of inappropriate        antidiuretic hormone secretion (SIADH).    -   5.43. Method 5.42 further comprising one or more of restriction        of dietary sodium, fluid and/or alcohol; and/or administration        of one or more diuretics, vasopressin receptor antagonists,        angiotensin converting enzyme (ACE) inhibitors, aldosterone        inhibitors, angiotensin receptor blockers (ARBs),        beta-adrenergic antagonists (beta-blockers), and/or digoxin.    -   5.44. Any of Method 5, et seq. wherein the compound of Formula I        or formula 1a inhibits aquaporin activity, e.g., AQP2 and/or        AQP4 activity, by at least 50% at concentrations of 10        micromolar or less, for example inhibits APQ2 and/or AQP4        activity by at least 50% at concentrations of 10 micromolar or        less in an aquaporin-mediated cell volume change assay, e.g., is        active in any of the assays of Method 10, et seq. infra    -   5.45. Any of Method 5, et seq. wherein the duration of treatment        with the phenylbenzamide is less than 21 days, e.g., less than 2        weeks, e.g., one week or less.    -   5.46. Any of Method 5, et seq. wherein the niclosamide or the        compound of Formula I or formula 1a is administered orally.    -   5.47. Any of Method 5, et seq. wherein the niclosamide or the        compound of Formula I or formula 1a is administered        parenterally.    -   5.48. Method 5.47 wherein the niclosamide or the compound of        Formula I or formula 1a is administered intravenously.    -   5.49. Any of Method 5, et seq. wherein the patient is human.    -   5.50. Any of Method 5, et seq. wherein the onset of action of        any of the compounds identified in Methods 1-1.28 is fairly        rapid.

In a further embodiment, the invention provides a method (Method 6) ofinhibiting an aquaporin comprising contacting the aquaporin with aneffective amount of a phenylbenzamide, e.g. niclosamide or a compound ofFormula I or formula 1a as hereinbefore described, e.g., any of thecompounds identified in Method 1-1.28 above, for example

6.

-   -   6.1. Method 6 wherein the aquaporin is inhibited in vivo.    -   6.2. Method 6 wherein the aquaporin is inhibited in vitro.    -   6.3. Any of Methods 6, et seq. wherein the aquaporin is AQP4.    -   6.4. Any of Method 6, et seq. wherein the aquaporin is AQP2.    -   6.5. Any of Method 6, et seq. wherein the compound of Formula I        or formula 1a inhibits aquaporin activity, e.g., AQP2 and/or        AQP4 activity, by at least 50% at concentrations of 10        micromolar or less, for example inhibits APQ2 and/or AQP4        activity by at least 50% at concentrations of 10 micromolar or        less in an aquaporin-mediated cell volume change assay, e.g., is        active in any of the assays of Method 10, et seq. infra    -   6.6. Method 6.1 wherein the niclosamide or the compound of        Formula I or formula 1a is administered orally.    -   6.7. Method 6.1 wherein the niclosamide or the compound of        Formula I or formula 1a is administered parenterally.    -   6.8. Method of 6.7 wherein the niclosamide or the compound of        Formula I or formula 1a is administered intravenously.

In a further embodiment, the invention provides a method (Method 7) toinhibit an aquaporin in a patient suffering from a disease or conditionmediated by an aquaporin comprising administering an effective amount ofa phenylbenzamide, e.g. niclosamide or a compound of Formula T orformula 1a as hereinbefore described, e.g., any of the compoundsidentified in Method 1-1.28 above, to inhibit the aquaporin.

7.

-   -   7.1. Method 7 wherein the aquaporin is AQP4.    -   7.2. Method 7 or 7.1 wherein the condition to be treated or        controlled is selected from edema, e.g. edema of the brain or        spinal cord, e.g., cerebral edema, e.g. cerebral edema        consequent to head trauma, ischemic stroke, glioma, meningitis,        acute mountain sickness, epileptic seizure, infection, metabolic        disorder, water intoxication, hepatic failure, hepatic        encephalopathy, or diabetic ketoacidosis or, e.g., spinal cord        edema, e.g., spinal cord edema consequent to spinal cord trauma,        e.g., spinal cord compression.    -   7.3. Method 7.2 further comprising a treatment selected from one        or more of the following: optimal head and neck positioning to        facilitate venous outflow, e.g. head elevation 30°; avoidance of        dehydration; systemic hypotension; maintenance of normothermia        or hypothermia; aggressive measures; osmotherapy, e.g., using        mannitol or hypertonic saline; hyperventilation; therapeutic        pressor therapy to enhance cerebral perfusion; administration of        barbiturates to reduce of cerebral metabolism (CMO₂);        hemicraniectomy; administration of aspirin; administration of        amantadine; intravenous thrombolysis (e.g. using rtPA);        mechanical clot removal; angioplasty; and/or stents.    -   7.4. Any of Method 7, et seq. wherein the patient is at elevated        risk of cerebral edema, e.g., due to head trauma, ischemic        stroke, glioma, meningitis, acute mountain sickness, epileptic        seizure, infection, metabolic disorder, water intoxication,        hepatic failure, hepatic encephalopathy, or diabetic        ketoacidosis.    -   7.5. Any of Method 7, et seq. wherein the patient has suffered a        stroke, head injury, or spinal injury.    -   7.6. Any of Method 7, et seq. wherein the patient has suffered a        stroke, head injury or spinal injury within 12 hours, e.g.        within 6 hours, preferably within 3 hours of commencing        treatment.    -   7.7. Any of Method 7, et seq. wherein the patient is at elevated        risk of suffering a stroke, head injury or spinal injury, e.g.,        in combat or in an athletic competition.    -   7.8. Any of Method 7, et seq. wherein the patient already has        cerebral edema.    -   7.9. Any of Method 7, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a stroke        or a traumatic brain injury.    -   7.10. Any of Method 7, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a middle        cerebral artery stroke.    -   7.11. Any of Method 7, et seq. wherein the condition to be        treated or controlled is cerebral edema consequent to a closed        head trauma.    -   7.12. Any of Methods 7-7.4 wherein the condition to be treated        or controlled is cerebral edema consequent to an epileptic        seizure.    -   7.13. Any of Methods 7-7.4 wherein the condition to be treated        or controlled is cerebral edema consequent an infection.    -   7.14. Any of Methods 7-7.4 wherein the condition to be treated        or controlled is cerebral edema consequent to a metabolic        disorder.    -   7.15. Any of Methods 7-7.4 wherein the condition to be treated        or controlled is cerebral edema consequent to glioma.    -   7.16. Any of Methods 7-7.4 wherein the condition to be treated        or controlled is cerebral edema consequent to meningitis, acute        mountain sickness, or water intoxication.    -   7.17. Any of Methods 7-7.4 wherein the condition to be treated        or controlled is cerebral edema consequent to hepatic failure,        hepatic encephalopathy, or diabetic ketoacidosis.    -   7.18. Any of Methods 7-7.3 wherein the condition to be treated        or controlled is cerebral edema consequent to an abscess.    -   7.19. Any of Methods 7-7.3 wherein the condition to be treated        or controlled is cerebral edema consequent to eclampsia.    -   7.20. Any of Methods 7-7.3 wherein the condition to be treated        or controlled is cerebral edema consequent to Creutzfeldt-Jakob        disease.    -   7.21. Any of Methods 7-7.3 wherein the condition to be treated        or controlled is cerebral edema consequent to lupus cerebritis.    -   7.22. Any of Methods 7-7.3 wherein the condition to be treated        or controlled is edema consequent to hypoxia, e.g., general        systemic hypoxia, e.g., hypoxia caused by an interruption of        blood perfusion, for example wherein the edema is cerebral edema        consequent to hypoxia caused by cardiac arrest, stroke, or other        interruption of blood perfusion to the brain, or wherein the        edema is cardiac edema consequent to cardiac ischemia or other        interruption of blood flow to the heart.    -   7.23. Any of Methods 7-7.3 wherein the condition to be treated        or controlled is cerebral and/or optical nerve edema consequent        to microgravity and/or radiation exposure, e.g., exposure from        space flight or from working with radioactive materials or from        working in radioactive areas.    -   7.24. Any of Methods 7-7.3 wherein the condition to be treated        or controlled is cerebral edema consequent to invasive central        nervous system procedures, e.g., neurosurgery, endovascular clot        removal, spinal tap, aneurysm repair, or deep brain stimulation.    -   7.25. Method 7.23 or 7.24 wherein the patient is at elevated        risk of edema, e.g., due to microgravity and/or radiation        exposure, neurosurgery, endovascular clot removal, spinal tap,        aneurysm repair, or deep brain stimulation.    -   7.26. Method 7.23 or 7.24 wherein the patient already has edema.    -   7.27. Any of Methods 7, et seq. wherein the edema is cytotoxic        cerebral edema or is primarily cytotoxic cerebral edema.    -   7.28. Any of Methods 7-7.17 or 7.22 wherein the edema is        cytotoxic cerebral edema or is primarily cytotoxic cerebral        edema.    -   7.29. Any of Methods 7-7.2 wherein the condition to be treated        or controlled is spinal cord edema, e.g., spinal cord edema        consequent to spinal cord trauma, e.g., spinal cord compression.    -   7.30. Method 7.29 wherein the condition to be treated or        controlled is spinal cord edema consequent to spinal cord        compression.    -   7.31. Any of Methods 7-7.2 wherein the condition to be treated        or controlled is retinal edema.    -   7.32. Method 7 or 7.1 wherein the condition to be treated or        controlled is epilepsy.    -   7.33. Method 7 or 7.1 wherein the condition to be treated or        controlled is retinal ischemia or other diseases of the eye        associated with abnormalities in intraocular pressure and/or        tissue hydration.    -   7.34. Method 7 or 7.1 wherein the condition to be treated or        controlled is myocardial ischemia.    -   7.35. Method 7 or 7.1 wherein the condition to be treated or        controlled is myocardial ischemia/reperfusion injury.    -   7.36. Method 7 or 7.1 wherein the condition to be treated or        controlled is myocardial infarction.    -   7.37. Method 7 or 7.1 wherein the condition to be treated or        controlled is myocardial hypoxia.    -   7.38. Method 7 or 7.1 wherein the condition to be treated or        controlled is congestive heart failure.    -   7.39. Method 7 or 7.1 wherein the condition to be treated or        controlled is sepsis.    -   7.40. Method 7 or 7.1 wherein the condition to be treated or        controlled is a migraine.    -   7.41. Method 7 wherein the aquaporin is AQP2.    -   7.42. Method 7 or 7.41 wherein the condition to be treated is        hyponatremia or excessive fluid retention, e.g., consequent to        heart failure (HF), for example congestive heart failure, liver        cirrhosis, nephrotic disorder, or syndrome of inappropriate        antidiuretic hormone secretion (SIADH).    -   7.43. Method 7.42 further comprising one or more of restriction        of dietary sodium, fluid and/or alcohol; and/or administration        of one or more diuretics, vasopressin receptor antagonists,        angiotensin converting enzyme (ACE) inhibitors, aldosterone        inhibitors, angiotensin receptor blockers (ARBs),        beta-adrenergic antagonists (beta-blockers), and/or digoxin.    -   7.44. Any of Method 7, et seq. wherein the compound of Formula 1        or formula 1a inhibits aquaporin activity, e.g., AQP2 and/or        AQP4 activity, by at least 50% at concentrations of 10        micromolar or less, for example inhibits APQ2 and/or AQP4        activity by at least 50% at concentrations of 10 micromolar or        less in an aquaporin-mediated cell volume change assay, e.g., is        active in any of the assays of Method 10, et seq. infra    -   7.45. Any of Method 7, et seq. wherein the duration of treatment        with the phenylbenzamide is less than 21 days, e.g., less than 2        weeks, e.g., one week or less.    -   7.46. Any of Method 7, et seq. wherein the niclosamide or        compound of Formula I or formula 1a is administered orally.    -   7.47. Any of Method 7, et seq. wherein the niclosamide or        compound of Formula I or formula 1a is administered        parenterally.    -   7.48. Method 7.47 wherein the niclosamide or compound of Formula        I or formula 1a is administered intravenously.    -   7.49. Any of Method 7, et seq. wherein the patient is human.    -   7.50. Any of Method 7, et seq. wherein the onset of action of        any of the compounds identified in Methods 1-1.28 is fairly        rapid.

In a further embodiment, the invention provides a pharmaceuticalcomposition comprising a phenylbenzamide, e.g. niclosamide or a compoundof Formula I or formula 1a as hereinbefore described, e.g., any of thecompounds identified in Methods 1-1.28 above, for use (Use 8) to inhibitan aquaporin in a patient suffering from a disease or condition mediatedby the aquaporin. For example, for use in any of the foregoing methods.

8.

-   -   8.1. Use 8 wherein the aquaporin is AQP4.    -   8.2. Use 8 or 8.1 wherein the condition to be treated or        controlled is selected from edema, e.g. edema of the brain or        spinal cord, e.g., cerebral edema, e.g. cerebral edema        consequent to head trauma, ischemic stroke, glioma, meningitis,        acute mountain sickness, epileptic seizure, infection, metabolic        disorder, water intoxication, hepatic failure, hepatic        encephalopathy, or diabetic ketoacidosis or, e.g., spinal cord        edema, e.g., spinal cord edema consequent to spinal cord trauma,        e.g., spinal cord compression.    -   8.3. Use 8.2 further comprising a treatment selected from one or        more of the following: optimal head and neck positioning to        facilitate venous outflow, e.g. head elevation 30°; avoidance of        dehydration; systemic hypotension; maintenance of normothermia        or hypothermia; aggressive measures; osmotherapy, e.g., using        mannitol or hypertonic saline; hyperventilation; therapeutic        pressor therapy to enhance cerebral perfusion; administration of        barbiturates to reduce of cerebral metabolism (CMO₂);        hemicraniectomy; administration of aspirin; administration of        amantadine; intravenous thrombolysis (e.g. using rtPA);        mechanical clot removal; angioplasty; and/or stents.    -   8.4. Any of Use 8, et seq. wherein the patient is at elevated        risk of cerebral edema, e.g., due to head trauma, ischemic        stroke, glioma, meningitis, acute mountain sickness, epileptic        seizure, infection, metabolic disorder, water intoxication,        hepatic failure, hepatic encephalopathy, or diabetic        ketoacidosis.    -   8.5. Any of Use 8, et seq. wherein the patient has suffered a        stroke, head injury, or spinal injury.    -   8.6. Any of Use 8, et seq. wherein the patient has suffered a        stroke, head injury or spinal injury within 12 hours, e.g.        within 6 hours, preferably within 3 hours of commencing        treatment.    -   8.7. Any of Use 8, et seq. wherein the patient is at elevated        risk of suffering a stroke, head injury or spinal injury, e.g.,        in combat or in an athletic competition.    -   8.8. Any of Use 8, et seq. wherein the patient already has        cerebral edema.    -   8.9. Any of Use 8, et seq. wherein the condition to be treated        or controlled is cerebral edema consequent to a stroke or a        traumatic brain injury.    -   8.10. Any of Use 8, et seq. wherein the condition to be treated        or controlled is cerebral edema consequent to a middle cerebral        artery stroke.    -   8.11. Any of Use 8, et seq. wherein the condition to be treated        or controlled is cerebral edema consequent to a closed head        trauma.    -   8.12. Any of Uses 8-8.4 wherein the condition to be treated or        controlled is cerebral edema consequent to an epileptic seizure.    -   8.13. Any of Uses 8-8.4 wherein the condition to be treated or        controlled is cerebral edema consequent to an infection.    -   8.14. Any of Uses 8-8.4 wherein the condition to be treated or        controlled is cerebral edema consequent to a metabolic disorder.    -   8.15. Any of Uses 8-8.4 wherein the condition to be treated or        controlled is cerebral edema consequent to glioma.    -   8.16. Any of Uses 8-8.4 wherein the condition to be treated or        controlled is cerebral edema consequent to meningitis, acute        mountain sickness, or water intoxication.    -   8.17. Any of Uses 8-8.4 wherein the condition to be treated or        controlled is cerebral edema consequent to hepatic failure,        hepatic encephalopathy, or diabetic ketoacidosis.    -   8.18. Any of the Uses 8-8.3 wherein the condition to be treated        or controlled is cerebral edema consequent to an abscess.    -   8.19. Any of the Uses 8-8.3 wherein the condition to be treated        or controlled is cerebral edema consequent to eclampsia.    -   8.20. Any of Uses 8-8.3 wherein the condition to be treated or        controlled is cerebral edema consequent to Creutzfeldt-Jakob        disease.    -   8.21. Any of Uses 8-8.3 wherein the condition to be treated or        controlled is cerebral edema consequent lupus cerebritis.    -   8.22. Any of Uses 8-8.3 wherein the condition to be treated or        controlled is edema consequent to hypoxia, e.g., general        systemic hypoxia, e.g., hypoxia caused by an interruption of        blood perfusion, for example wherein the edema is cerebral edema        consequent to hypoxia caused by cardiac arrest, stroke, or other        interruption of blood perfusion to the brain, or wherein the        edema is cardiac edema consequent to cardiac ischemia or other        interruption of blood flow to the heart.    -   8.23. Any of Uses 8-8.3 wherein the condition to be treated or        controlled is cerebral and/or optic nerve edema consequent to        microgravity and/or radiation exposure, e.g., exposure from        space flight or from working with radioactive materials or from        working in radioactive areas.    -   8.24. Any of Uses 8-8.3 wherein the condition to be treated or        controlled is cerebral edema consequent to invasive central        nervous system procedures, e.g., neurosurgery, endovascular clot        removal, spinal tap, aneurysm repair, or deep brain stimulation.    -   8.25. Use 8.23 or 8.24 wherein the patient is at elevated risk        of edema, e.g., due to microgravity exposure and/or radiation,        neurosurgery, endovascular clot removal, spinal tap, aneurysm        repair, or deep brain stimulation.    -   8.26. Use 8.23 or 8.24 wherein the patient already has edema.    -   8.27. Any of Uses 8, et seq. wherein the edema is cytotoxic        cerebral edema or is primarily cytotoxic cerebral edema.    -   8.28. Any of Uses 8-8.17 or 8.22 wherein the edema is cytotoxic        cerebral edema or is primarily cytotoxic cerebral edema.    -   8.29. Any of Uses 8-8.2 wherein the condition to be treated or        controlled is spinal cord edema, e.g., spinal cord edema        consequent to spinal cord trauma, e.g., spinal cord compression.    -   8.30. Use 8.29 wherein the condition to be treated or controlled        is spinal cord edema consequent to spinal cord compression.    -   8.31. Any of Uses 8-8.2 wherein the condition to be treated or        controlled is retinal edema.    -   8.32. Use 8 or 8.1 wherein the condition to be treated or        controlled is epilepsy.    -   8.33. Use 8 or 8.1 wherein the condition to be treated or        controlled is retinal ischemia or other diseases of the eye        associated with abnormalities in intraocular pressure and/or        tissue hydration.    -   8.34. Use 8 or 8.1 wherein the condition to be treated or        controlled is myocardial ischemia.    -   8.35. Use 8 or 8.1 wherein the condition to be treated or        controlled is myocardial ischemia/reperfusion injury.    -   8.36. Use 8 or 8.1 wherein the condition to be treated or        controlled is myocardial infarction.    -   8.37. Use 8 or 8.1 wherein the condition to be treated or        controlled is myocardial hypoxia.    -   8.38. Use 8 or 8.1 wherein the condition to be treated or        controlled is congestive heart failure.    -   8.39. Use 8 or 8.1 wherein the condition to be treated or        controlled is sepsis.    -   8.40. Use 8 or 8.1 wherein the condition to be treated or        controlled is a migraine.    -   8.41. Use 8 wherein the aquaporin is AQP2.    -   8.42. Use 8 or 8.41 wherein the condition to be treated is        hyponatremia or excessive fluid retention, e.g., consequent to        heart failure (HF), for example congestive heart failure, liver        cirrhosis, nephrotic disorder, or syndrome of inappropriate        antidiuretic hormone secretion (SIADH).    -   8.43. Use 8.42 further comprising one or more of restriction of        dietary sodium, fluid and/or alcohol; and/or administration of        one or more diuretics, vasopressin receptor antagonists,        angiotensin converting enzyme (ACE) inhibitors, aldosterone        inhibitors, angiotensin receptor blockers (ARBs),        beta-adrenergic antagonists (beta-blockers), and/or digoxin.    -   8.44. Any of Use 8, et seq. wherein the compound of Formula I or        formula 1a inhibits aquaporin activity, e.g., AQP2 and/or AQP4        activity, by at least 50% at concentrations of 10 micromolar or        less, for example inhibits APQ2 and/or AQP4 activity by at least        50% at concentrations of 10 micromolar or less in an        aquaporin-mediated cell volume change assay, e.g., is active in        any of the assays of Method 10, et seq. infra    -   8.45. Any of Use 8, et seq. wherein the duration of treatment        with the phenylbenzamide is less than 21 days, e.g., less than 2        weeks, e.g., one week or less.    -   8.46. Any of Use 8, et seq. wherein the pharmaceutical        composition is administered orally.    -   8.47. Any of Use 8, et seq. wherein the pharmaceutical        composition is administered parenterally.    -   8.48. Use 8.47 wherein the pharmaceutical composition is        administered intravenously.    -   8.49. Any of Use 8, et seq. wherein the patient is human.    -   8.50. Any of Use 8, et seq. wherein the onset of action of the        pharmaceutical composition is fairly rapid.

In a further embodiment, the invention provides use (Use 9) of aphenylbenzamide, e.g. niclosamide or a compound of Formula I or formula1a as hereinbefore described, e.g., any of the compounds identified inMethods 1-1.28 above, in the manufacture of a medicament for treating orcontrolling a disease or condition mediated by an aquaporin wherein themedicament comprises the phenylbenzamide in an amount effective toinhibit the aquaporin. For example, for use in any of the foregoingmethods.

9.

-   -   9.1. Use 9 wherein the aquaporin is AQP4.    -   9.2. Any of Use 9 or 9.1 wherein the condition to be treated or        controlled is selected from edema, e.g. edema of the brain or        spinal cord, e.g., cerebral edema, e.g. cerebral edema        consequent to head trauma, ischemic stroke, glioma, meningitis,        acute mountain sickness, epileptic seizure, infection, metabolic        disorder, water intoxication, hepatic failure, hepatic        encephalopathy, or diabetic ketoacidosis or, e.g., spinal cord        edema, e.g., spinal cord edema consequent to spinal cord trauma,        e.g., spinal cord compression.    -   9.3. Use 9.2 further comprising a treatment selected from one or        more of the following: optimal head and neck positioning to        facilitate venous outflow, e.g. head elevation 30°; avoidance of        dehydration; systemic hypotension; maintenance of normothermia        or hypothermia; aggressive measures; osmotherapy, e.g., using        mannitol or hypertonic saline; hyperventilation; therapeutic        pressor therapy to enhance cerebral perfusion; administration of        barbiturates to reduce of cerebral metabolism (CMO₂);        hemicraniectomy; administration of aspirin; administration of        amantadine; intravenous thrombolysis (e.g. using rtPA);        mechanical clot removal; angioplasty; and/or stents.    -   9.4. Any of Use 9, et seq. wherein the patient is at elevated        risk of cerebral edema, e.g., due to head trauma, ischemic        stroke, glioma, meningitis, acute mountain sickness, epileptic        seizure, infection, metabolic disorder, water intoxication,        hepatic failure, hepatic encephalopathy, or diabetic        ketoacidosis.    -   9.5. Any of Use 9, et seq. wherein the patient has suffered a        stroke, head injury, or spinal injury.    -   9.6. Any of Use 9, et seq. wherein the patient has suffered a        stroke, head injury or spinal injury within 12 hours, e.g.        within 6 hours, preferably within 3 hours of commencing        treatment.    -   9.7. Any of Use 9, et seq. wherein the patient is at elevated        risk of suffering a stroke, head injury or spinal injury, e.g.,        in combat or in an athletic competition.    -   9.8. Any of Use 9, et seq. wherein the patient already has        cerebral edema.    -   9.9. Any of Use 9, et seq. wherein the condition to be treated        or controlled is cerebral edema consequent to a stroke or a        traumatic brain injury.    -   9.10. Any of Use 9, et seq. wherein the condition to be treated        or controlled is cerebral edema consequent to a middle cerebral        artery stroke.    -   9.11. Any of Use 9, et seq. wherein the condition to be treated        or controlled is cerebral edema consequent to a closed head        trauma.    -   9.12. Any of Uses 9-9.4 wherein the condition to be treated or        controlled is cerebral edema consequent to an epileptic seizure.    -   9.13. Any of Uses 9-9.4 wherein the condition to be treated or        controlled is cerebral edema consequent to an infection.    -   9.14. Any of Uses 9-9.4 wherein the condition to be treated or        controlled is cerebral edema consequent to a metabolic disorder.    -   9.15. Any of Uses 9-9.4 wherein the condition to be treated or        controlled is cerebral edema consequent to glioma.    -   9.16. Any of Uses 9-9.4 wherein the condition to be treated or        controlled is cerebral edema consequent to meningitis, acute        mountain sickness, or water intoxication.    -   9.17. Any of Uses 9-9.4 wherein the condition to be treated or        controlled is cerebral edema consequent to hepatic failure,        hepatic encephalopathy, or diabetic ketoacidosis.    -   9.18. Any of Uses 9-9.3 wherein the condition to be treated or        controlled is cerebral edema consequent to an abscess.    -   9.19. Any of Uses 9-9.3 wherein the condition to be treated or        controlled is cerebral edema consequent to eclampsia.    -   9.20. Any of Uses 9-9.3 wherein the condition to be treated or        controlled is cerebral edema consequent to Creutzfeldt-Jakob        disease.    -   9.21. Any of Uses 9-9.3 wherein the condition to be treated or        controlled is cerebral edema consequent to lupus cerebritis.    -   9.22. Any of Uses 9-9.3 wherein the condition to be treated or        controlled is edema consequent to hypoxia, e.g., general        systemic hypoxia, e.g., hypoxia caused by an interruption of        blood perfusion, for example wherein the edema is cerebral edema        consequent to hypoxia caused by cardiac arrest, stroke, or other        interruption of blood perfusion to the brain, or wherein the        edema is cardiac edema consequent to cardiac ischemia or other        interruption of blood flow to the heart.    -   9.23. Any of Uses 9-9.3 wherein the condition to be treated or        controlled is cerebral and/or optic nerve edema consequent to        microgravity and/or radiation exposure, e.g., exposure from        space flight or from working with radioactive materials or from        working in radioactive areas.    -   9.24. Any of Uses 9-9.3 wherein the condition to be treated or        controlled is cerebral edema consequent to invasive central        nervous system procedures, e.g., neurosurgery, endovascular clot        removal, spinal tap, aneurysm repair, or deep brain stimulation.    -   9.25. Use 9.24 or 9.25 wherein the patient is at elevated risk        of edema, e.g., due to microgravity and/or radiation exposure,        neurosurgery, endovascular clot removal, spinal tap, aneurysm        repair, or deep brain stimulation.    -   9.26. Use 9.24 or 9.25 wherein the patient already has edema.    -   9.27. Any of Uses 9, et seq. wherein the edema is cytotoxic        cerebral edema or is primarily cytotoxic cerebral edema.    -   9.28. Any of Uses 9-9.17 or 9.22 wherein the edema is cytotoxic        cerebral edema or is primarily cytotoxic cerebral edema.    -   9.29. Any of Uses 9-9.2 wherein the condition to be treated or        controlled is spinal cord edema, e.g., spinal cord edema        consequent to spinal cord trauma, e.g., spinal cord compression.    -   9.30. Use 9.29 wherein the condition to be treated or controlled        is spinal cord edema consequent to spinal cord compression.    -   9.31. Any of Uses 9-9.2 wherein the condition to be treated or        controlled is retinal edema.    -   9.32. Use 9 or 9.1 wherein the condition to be treated or        controlled is epilepsy.    -   9.33. Use 9 or 9.1 wherein the condition to be treated or        controlled is retinal ischemia or other diseases of the eye        associated with abnormalities in intraocular pressure and/or        tissue hydration.    -   9.34. Use 9 or 9.1 wherein the condition to be treated or        controlled is myocardial ischemia.    -   9.35. Use 9 or 9.1 wherein the condition to be treated or        controlled is myocardial ischemia/reperfusion injury.    -   9.36. Use 9 or 9.1 wherein the condition to be treated or        controlled is myocardial infarction.    -   9.37. Use 9 or 9.1 wherein the condition to be treated or        controlled is myocardial hypoxia.    -   9.38. Use 9 or 9.1 wherein the condition to be treated or        controlled is congestive heart failure.    -   9.39. Use 9 or 9.1 wherein the condition to be treated or        controlled is sepsis.    -   9.40. Use 9 or 9.1 wherein the condition to be treated or        controlled is a migraine.    -   9.41. Use 9 wherein the aquaporin is AQP2.    -   9.42. Use 9 or 9.41 wherein the condition to be treated is        hyponatremia or excessive fluid retention, e.g., consequent to        heart failure (HF), for example congestive heart failure, liver        cirrhosis, nephrotic disorder, or syndrome of inappropriate        antidiuretic hormone secretion (SIADH).    -   9.43. Use 9.42 further comprising one or more of restriction of        dietary sodium, fluid and/or alcohol; and/or administration of        one or more diuretics, vasopressin receptor antagonists,        angiotensin converting enzyme (ACE) inhibitors, aldosterone        inhibitors, angiotensin receptor blockers (ARBs),        beta-adrenergic antagonists (beta-blockers), and/or digoxin.    -   9.44. Any of Use 9, et seq. wherein the compound of Formula I or        formula 1a inhibits aquaporin activity, e.g., AQP2 and/or AQP4        activity, by at least 50% at concentrations of 10 micromolar or        less, for example inhibits APQ2 and/or AQP4 activity by at least        50% at concentrations of 10 micromolar or less in an        aquaporin-mediated cell volume change assay, e.g., is active in        any of the assays of Method 10, et seq. infra    -   9.45. Any of Use 9, et seq. wherein the duration of treatment        with the phenylbenzamide is less than 21 days, e.g., less than 2        weeks, e.g., one week or less.    -   9.46. Any of Use 9, et seq. wherein the medicament is formulated        for oral administration.    -   9.47. Any of Use 9, et seq. wherein the medicament is formulated        for parenteral administration.    -   9.48. Use 9.47 wherein the medicament is formulated for        intravenous administration.

A dose or method of administration of the dose of the present inventionis not particularly limited. Dosages employed in practicing the presentinvention will of course vary depending, e.g. on the particular diseaseor condition to be treated, the particular compound used, the mode ofadministration, and the therapy desired. The compounds may beadministered by any suitable route, including orally, parenterally,transdermally, or by inhalation. In stroke or other severelydebilitating diseases or conditions, for example where the patient maybe unconscious or unable to swallow, an IV infusion or IV bolus may bepreferred. In general, satisfactory results, e.g. for the treatment ofdiseases as hereinbefore set forth are indicated to be obtained on oraladministration at dosages of the order from about 0.01 to 15.0 mg/kg. Inlarger mammals, for example humans, an indicated daily dosage for oraladministration will accordingly be in the range of from about 0.75 to1000 mg per day, conveniently administered once, or in divided doses 2to 3 times, daily or in sustained release form. Unit dosage forms fororal administration thus for example may comprise from about 0.2 to 75or 150 mg, e.g. from about 0.2 or 2.0 to 50, 75, 100, 125, 150 or 200 mgof a Compound of the Invention, together with a pharmaceuticallyacceptable diluent or carrier therefor. When the medicament is used viainjection (subcutaneously, intramuscularly or intravenously) the dosemay be 0.25 to 500 mg per day by bolus or if IV by bolus or infusion.

Pharmaceutical compositions comprising compounds of Formula I or formula1a may be prepared using conventional diluents or excipients andtechniques known in the galenic art. Thus oral dosage forms may includetablets, capsules, solutions, suspensions and the like.

Methods of making and formulating compounds of Formula I or formula 1aare set forth in US 2010/0274051 A1, U.S. Pat. No. 7,700,655, and inU.S. Pat. No. 7,626,042, each incorporated herein by reference.

In a further embodiment, the invention provides a method, e.g., Method10, for identification of specific aquaporin inhibitors, comprisingmeasuring the response of an aquaporin-expressing cell population versusa control cell population to a hypertonic or hypotonic solution in thepresence or absence of a test compound. For example the inventionprovides, e.g.,

10.

-   -   10.1. Method 10 wherein the aquaporin-expressing cell population        expresses AQP2 or AQP4.    -   10.2. Any of Method 10 or 10.1 wherein the cells are mammalian,        e.g., transgenic CHO cells.    -   10.3. Any of Method 10, et seq. wherein the control cells        express a transgenic transmembrane protein other than an        aquaporin, e.g., CD81.    -   10.4. Any of Method 10, et seq. wherein the cells are exposed to        a hypotonic environment for a period of time and at a        concentration which causes most of the aquaporin-expressing cell        population to burst in the absence of test compound, but not the        control cell population, e.g. 3-8 minutes in water.    -   10.5. Method 10.4 wherein cell bursting is measured by a        fluorescent signal that is produced by viable cells but not by        burst cells, e.g., conversion of acetoxymethyl calcein        (calcein-AM) to the fluorescent dye calcein.    -   10.6. Method 10.4 or 10.5 wherein the cells are exposed to a        hypotonic environment, e.g., deionized water, for a period of        3-8 minutes, and then returned to normotonic environment (e.g.        ca. 300 mOSM) then the proportion of viable cells is measured.    -   10.7. Any of Method 10, et seq. wherein the measurement of        viability is the ability of the cells to convert acetoxymethyl        calcein (calcein-AM) to the fluorescent dye calcein.    -   10.8. Any of the foregoing Methods 10.4, et seq. wherein a        compound of Formula I as hereinbefore described, e.g., of        formula 1a, is used as a positive control, which inhibits the        aquaporin-expressing cell population from bursting in a        hypotonic environment.    -   10.9. Any of the foregoing Methods 10.4-10.8 wherein a test        compound is identified as having aquaporin-inhibitory activity        when the aquaporin-expressing cell population is identified        viable, e.g., by the ability of the cells to convert        acetoxymethyl calcein (calcein-AM) to the fluorescent dye        calcein, following an exposure in a presence of test compound to        a hypotonic environment that renders the aquaporin-expressing        cell population non-viable in the absence of test compound.    -   10.10. Any of Method 10 or 10.1-10.3 wherein the cells are        exposed to a hypertonic environment for a period of time and at        a concentration sufficient to cause the aquaporin-expressing        cell population to shrink in the absence of test compound, e.g.        3-8 minutes at ca. 530 mOsm.    -   10.11. Method 10.9 wherein the shrinkage of the cells is        measured by light scattering.    -   10.12. Any of Method 10.10, et seq. wherein the hypertonic        environment is approximately 530 mOsm and the normotonic        environment is approximately 300 mOsm.    -   10.13. Any of the foregoing Methods 10.10, et seq. wherein a        compound of Formula I as hereinbefore described, e.g., of        formula 1a, is used as a positive control, which inhibits the        aquaporin-expressing cell population from shrinking in a        hypertonic environment.    -   10.14. Any of the foregoing Methods 10.10, et seq. wherein a        test compound is identified as inhibiting aquaporin activity by        inhibiting the aquaporin-expressing cell population from        shrinking in a hypertonic environment.    -   10.15. Any of the foregoing methods wherein a test compound is        identified as inhibiting aquaporin activity.    -   10.16. Any of Method 10, et seq. wherein the        aquaporin-expressing cell population expresses AQP2.    -   10.17. Any of Method 10, et seq. wherein the        aquaporin-expressing cell population expresses AQP4.    -   10.18. Any of Method 10, et seq. wherein the test compound is a        phenylbenzamide, e.g., of Formula I as hereinbefore described,        e.g., of formula 1a.

As used throughout, ranges are used as shorthand for describing each andevery value that is within the range. Any value within the range can beselected as the terminus of the range. In addition, all references citedherein are hereby incorporated by referenced in their entireties. In theevent of a conflict in a definition in the present disclosure and thatof a cited reference, the present disclosure controls.

Unless otherwise specified, all percentages and amounts expressed hereinand elsewhere in the specification should be understood to refer topercentages by weight. The amounts given are based on the active weightof the material.

EXAMPLE 1 Phenylbenzamide-AQP Structure-Activity Relationship

Structure activity relationships (SARs) are determined by assayinganalogues of selected hits to guide chemistry for the preparation of newmolecules to be tested for improved potency. For this iterative processwe use a quantitative kinetic assay—the Aquaporin-Mediated Cell VolumeChange Assa—in a 96-well multiplate reader. It detects changes in lightscattering by a monolayer of CHO cells expressing the desired AQP asthey shrink when exposed to hypertonic solution (300 mOsm→530 mOsm).FIG. 1 depicts the aquaporin-mediated cell volume change assay with AQP4expressing cells (FIG. 1A) and AQP2 expressing cells (FIG. 1B). Thecells expressing aquaporins shrink more rapidly than control cells, dueto enhanced water flow, which shrinkage can be inhibited by a compoundthat inhibits the aquaporin.

In FIG. 1, aquaporin-expressing cells are shown in the presence of DMSO(triangles) or in the presence of the test compound (here, Compound 3)at 10 μM (squares), along with CHO-CD81 expressing control cells in thepresence of DMSO (diamonds). Each curve represents an average of 16wells in the 96-well plate.

In FIG. 1A, when the AQP4b cells treated with DMSO are exposed tohypertonic shock, the cells show rapid shrinking, giving a rise in lightscattering (increasing relative change in absorbance, Abs/Abs₀) followedby a decay as cells detach from the plate. The CHO-AQP4b cell line showsa 4.5-fold increase in the rate of shrinking compared to CHO-CD81control cells (fitted to a double exponential model). CHO-AQP4b cellstreated with the Compound 3 analogue at 1004 (squares) show a slowerrate of shrinking (55% inhibition) as seen by characteristic ‘unbending’of the light scattering curve. Similarly, FIG. 1B depicts an experimentcomparing CHO-AQP2 treated with DMSO or with Compound 3 at 10 μM.Aquaporin-2 has a lower intrinsic water permeability than AQP4 asobserved here. CHO-AQP2 cell lines treated with DMSO (FIG. 1B,triangles) show a 1.7-fold increase in the rate of shrinking compared toCHO-CD81 control cells (diamonds) also treated with DMSO (fitted to adouble exponential model) (FIG. 1B). CHO-AQP2 cells treated withCompound 3 at 10 μM (squares) show a slower rate of shrinking (81%inhibition), when comparing the relative change in Abs (Abs/Abs₀) (FIG.1B).

The data indicates that in this assay, Compound 3 is capable ofsignificantly inhibiting AQP2 and AQP4 activity, e.g. by greater than50%, at concentrations of 10

EXAMPLE 2 Aquaporin Specificity of the Phenylbenzamide Compounds

The specificity of the compounds is tested against the most closelyrelated of the 13 known aquaporins: AQP1, AQP2, AQP5 and both splicevariants of AQP4 (A and B). A stable CHO cell line is created for eachof the above aquaporins and the inhibition of water permeability usingthe Aquaporin-Mediated Cell Volume Change Assay with 10 μM Compound 3 istested. Compound 3 inhibits AQP2 and 4, while it poorly inhibits AQP1and 5 (FIG. 2).

EXAMPLE 3 Direct Drug-Target Interactions Between Phenylbenzamides andAQP4

To support the mechanism of action by which phenylbenzamides directlyblock AQP4, we perform in vitro binding studies using purified AQP4b andCompound 4 radiolabeled with ³H. Using a Hummel-Dryer style assay, a gelfiltration column is equilibratrated with buffer containing detergent,to maintain solubility of AQP4b, and 1 μM [³H]-Compound 4. AQP4b isdiluted to 250 μM in this column buffer and incubated at RT for 30 min.The sample is then applied to the column, fractions collected and thepresence of [³H]-Compound 4 detected by liquid scintillation counting.FIG. 3 shows the elution profile of [³H]-Compound 4 from the gelfiltration column with the elution positions of tetrameric and monomericAQP4b indicated. The rise in [³H]-Compound 4 from a baseline value of 1μM represents binding to each of these proteins. Although no monomericAQP4b can be readily detected in our highly purified AQP4b byconventional means, this assay reveals the presence of a small, albietvanishing, amount of monomer. The relative affinities for Compound 4 are˜100 μM and less than 1 μM for tetramer and monomer, respectively. Thisassay shows relatively weak binding of Compound 4 to solubilized AQP4b;nevertheless, it clearly demonstrates that this phenylbenzamide directlyinteracts with AQP4b.

EXAMPLE 4 Pharmacological Proof-of-Concept

Mouse Water Toxicity Model—Survival Curves: The in vivo efficacies ofthe compounds are tested using the mouse water toxicity model, where amouse is injected with water at 20% of its body weight. Manley, G. T. etal. Aquaporin-4 deletion in mice reduces brain edema after acute waterintoxication and ischemic stroke. Nat Med 6, 159-163 (2000); Gullans, S.R. & Verbalis, J. G. Control of brain volume during hyperosmolar andhypoosmolar conditions. Annual Review of Medicine 44, 289-301 (1993).The resulting euvolemic hyponatremia rapidly leads to CE, making this apractical model to test an inhibitor of the CNS aquaporin, AQP4b.

The ability of mice to survive H₂O toxicity is determined in threeexperiments using 10-12 mice each (16-19 weak old male/female).Deionized water is prepared for injection with either 0.39 mg/kgphenylbenzamide (placebo) or 0.76 mg/kg with test compound. FIG. 4 showsthe combined results of these experiments (n=33 placebo, n=34 Compound1). Percent survival of the Compound 1 cohorts improves 3.2 fold and thetime to 50% survival for animals treated with Compound 1 is improved byroughly 52 min.

Mouse Water Toxicity Model—Brain Water Content: Compounds are alsotested for the ability to reduce CE in mice exposed to the water shockby examining brain water content. Mice are treated with a water bolus asdescribed above, then sacrificed at 30 minutes. Brain water content isassessed by fresh/dry weight ratio, i.e. the brains are weighedimmediately then desiccated in an oven at 100° C. for 24 h and dryweights measured (brain water percentage=100×dry weight/fresh weight).At 30 minutes, mice treated with 0.76 mg/kg and 7.6 mg/kg Compound 1show an 11.2% and 15.9% reduction in CE, respectively (FIG. 5).

Mouse Water Toxicity Model—Brain Volume by Magnetic Resonance Imaging(MRI): MRI is used to measure changes in brain volume in response towater shock, using the water toxicity model. As described for thesurvival and brain water content studies above, mice are injected, IP,with a water bolus alone or water bolus and test compound at 0.76 mg/kg,and changes in brain volume as detected by MRI are monitored. Mousebrain volumes are assessed using MRI scans collected with a 9.4T BrukerBiospec MRI scanner at the Case Center for Imaging Research at CaseWestern Reserve University. This imaging method is found to providesufficient contrast and resolution to sensitively detect changes intotal brain volume in the mouse water toxicity model for cerebral edema.High resolution T2-weighted sagittal scans (resolution=0.1 mm×0.1 mm×0.7mm) of the mouse head are obtained prior to water injection, 5.67 minpost water injection, and then every 5.2 minutes until the animalexpires from the water loading. Each scan contains twenty-five 0.7 mmcontiguous imaging slices of which 14-15 slices contain a portion of thebrain. The cross sectional area of the brain in each imaging slice ismeasured by manual region-of-interest selection using ImageJ Brainvolumes are then calculated for each scan by summing the individualcross sectional brain areas and multiplying by the slice thickness (0.7mm).

Treatment with Compound 1 at 0.76 mg/kg reduces the rate of CEdevelopment from 0.081 to 0.032 min⁻¹ (or 2.5-fold) fit to a singleexponential model (FIG. 6). Also, the extent of CE during the period ofobservation is reduced (FIG. 6). Moreover, plasma levels in the sameassay are found to range between 0.03-0.06 μg as determined by LC-MS/MS(performed at Lerner Center, Cleveland Clinic, Cleveland, Ohio) and aresufficient to show efficacy in this model for CE.

The brain volume by magnetic resonance imaging experiment is alsoconducted with phenylbenzamide (0.39 mg/kg) and Compound 4 (0.83 mg/kg).Compound 4 reduces the rate of CE development from 0.081 to 0.022 min⁻¹(Table 1). Phenylbenzamide fails to show reduction in the rate of CE inmice (Table 1).

TABLE 1 Efficacy of compounds on CE formation in the mouse watertoxicity model AQP Inhibition Cerebral Edema Compound Cell-Based Assay(%) Rate by MRI (min⁻¹) No Drug 0 0.081 Compound 1 47.9 0.032Phenylbenzamide 4.5 0.096 Compound 4 38.9 0.022For no drug and Compound 1, n=14 mice each. For phenylbenzamide andCompound 4, n=12 mice each.

EXAMPLE 5 High Throughput Screening Assay

Under hypotonic shock, both untransfected cells and cells expressing anunrelated transmembrane protein (CD81, at levels equivalent to AQP4b)swell slowly but remain intact. These observations are used to developour high-throughput screening assay (HTS).

After hypotonic shock in a 384 well plate format, we return osmolalityto normal (300 mOSM) by adding 2× concentrated phosphate buffered salinesupplemented to 2 μM with a nonfluorescent acetoxymethyl derivative ofcalcein (calcein-AM) to each well. Intact cells take up calcein-AM andconvert it to the fluorescent dye calcein giving a quantitative measureof the remaining intact cells. Burst cells do not convert the precursorto the dye. Water uptake by AQP4-expressing cells is relatively rapid,with most test cells bursting within 4 min of hypotonic shock, whereasmost cells expressing CD81 remain viable after 8 min. Intracellularconversion of calcein-AM provides a strong and easily detectable signalat 535 nM in our assay (FIG. 7).

Calcein Fluorescence End-Point Assay:

Cells are seeded 24 hr before assay to reach 100% confluence. Culturemedium was replaced with H₂O for 5:30 min (osmotic shock). Osmolality isthen normalized with the addition of 2×PBS plus 2 μM calcein-AM. Cellsare then incubated at 37° C. for an additional 30 min and fluorescencemeasured on a plate-reader. Rows 1-22 are seeded with CHO-AQP4 cells,and rows 23-24, with CHO-CD81 cells (384 well plate). Note, all plateedges are discarded. Relative Fluorescence Intensity is calculated asthe fluorescence intensity (FI) of each well divided by the mean FI ofAQP4 cells treated with DMSO (control). Criteria for a successful assay:coefficients of variation (CVs) <15%, and Z-factors >0.5. Statisticalanalysis shows that 5.5 min of osmotic shock provides the optimalsignal-to-noise ratio.

TABLE 2 Statistics for endpoint ‘calcein’ assay in FIG. 7; 5:30 min timepoint shown: Mean StDev CV Z′ S/B AQP4 581618 66311 11% 0.629 5.0 CD812910106 221240  8%

As will be observed, the signal for the CD81 cells is ca. 5× higher thanthe signal for the APQ4 cells, because by 5.5 mins, most of the AQP4cells have burst, while most of the CD81 cells remain intact. Inhibitionof AQP4 would therefore be expected to provide a higher signal, morelike the CD81 cells.

This assay is applied in a pilot screen of the MicroSource GenPlus 960and the Maybridge Diversity™ 20 k libraries (approximately 21,000compounds tested, each compound at 10-20 μM).

From this assay, a specific chemical series is identified,phenylbenzamides, which represents 3 out of the top 234 hits.

Hits from the HTS are validated using the same assay using a differentplating arrangement. In FIG. 8, we show this validation assay used toexamine Compound 3. Cells are seeded in a 96 well multiplate format withthe plates edges omitted (lanes 1 and 24) and an entire column (n=16) isused to test the ability of a compound to block AQP4-mediated cellbursting upon H₂O shock. CHO cells expressing CD81 are seeded in lanes2-3 as a control, and CHO cells expressing AQP4, in lanes 4-23. Cellsare treated with 0.1% DMSO in 10% FBS, DMEM (even numbered columns) or10 μM Compound 1 (odd number columns) in 0.1% DMSO, 10% FBS, DMEM for 30minutes. The cells are shocked with H₂O for 5:30 minutes, thenosmolality returned to 300 mOSM in the presence of 1 μM calcein-AM, asdescribed above. The cells are incubated at 37° C. for 30 minutes andthe relative fluoresence measured (ex 495/em 535 nM) on a fluoresencemultiplate reader. The data in FIG. 8 represents the average relativefluoresence units (RFU±SEM, n=16).

EXAMPLE 6 Water Toxicity Model for CE: Intracranial Pressure (ICP)

ICP is monitored using a Samba 420 Sensor, pressure transducer, with aSamba 202 control unit (Harvard Apparatus, Holliston, Mass.). This ICPmonitoring system consists of a 0.42 mm silicon sensor element mountedon an optical fiber. A 20-gauge syringe needle is implanted through thecisterna magna to a depth of ˜1 cm. The needle then acts as a guide forinsertion of the Samba Sensor and the site of implantation and the openend of the needle are sealed with 100% silicone sealant. A baseline ICPreading is established followed by a water bolus IP injection (20%weight of animal) with or without Compound 1. ICP is monitored until theanimal expires from the water load.

Adjusting for the slight rise in ICP observed in the animals when theyare monitored without the water bolus injection (FIG. 9, No WaterToxicity), Compound 1 at 0.76 mg/kg reduces the relative rate of ICPrise by 36%, from 3.6×10⁻³ min⁻¹ to 2.3×10⁻³ min⁻¹ (n=6 mice/treatment,mean±SEM).

EXAMPLE 7 Conversion of Compound 5 to Compound 1

Plasma or serum levels of Compound 1 are measured by LC-MS/MS at theMass Spectrometry II Core facility at the Lerner Research Institute ofthe Cleveland Clinic Foundation. Measurements are taken at 15 minutesand 24 hours after a 10 mg/kg i.p. loading dose and 1 mg/ml at 8 μl/hmaintenance dose (delivered by an Alzet i.p. osmotic pump, Durect Corp.,Cupertino, Calif.) of Compound 5 (n=5 mice/time point, mean±SEM) (FIG.10). After initial processing to remove proteins (75% acetonitrileextraction), Compound 3 is introduced to improve quantitation usingmultiple reaction monitoring (MRM). Samples are analyzed by tandemLC-MS/MS using C18 reversed-phase chromatography and mass analysis witha triple-quadrapole mass spectrometer. The LC method is sufficient toseparate Compound 1 from Compound 3 and subsequent MRM gave reliablequantitation with a linear response from 0.004-0.4 ng of Compound 1 forits most abundant daughter ion. The dashed line in FIG. 10 is therelative effective plasma concentration of Compound 1 observed in themouse water toxicity model. Inclusion of an Alzet osmotic pump (DurectCorp., Cupertino, Calif.) containing Compound 5 in the peritoneum wassufficient, in conjunction with an initial loading dose, to sustainCompound 1 above the expected efficacious plasma concentration of 20ng/ml for 24 hours (FIG. 10).

The solubility of Compound 1 in water is 3.8 The solubility of Compound5 in water is 1 mg/ml. Initial experiments show rapid bioconversion ofCompound 5 to Compound 1 when added to mouse plasma in vitro. Less than5 minutes at 20° C. is sufficient to render Compound 5 undetectable. Inaddition, Compound 1 is undetectable in plasma samples taken from miceinjected IP with Compound 5. Instead, Compound 1 is detected at aconcentration consistent with good bioavailability and near-completeconversion of Compound 5. With compound 5, doses of 10 mg/kg and IPinjection volumes in saline (0.5 ml for a 30 g mouse), that give serumconcentrations of Compound 1 in excess of 400 ng/ml (FIG. 10) can beused. Key PK parameters for Compound 5 are: rate of absorption 0.12min⁻¹; rate of elimination 0.017 min⁻¹.

EXAMPLE 8 Animal Stroke Model

Most ischemic strokes (˜80%) occur in the region of the middle cerebralartery (MCA). To mimic this injury in mice, an intraluminal monofilamentmodel of middle cerebral artery occlusion (MCAo) is used. Occlusion isachieved by inserting a surgical filament into the external carotidartery (ECA) and threading it forward into the internal carotid artery(ICA) until the tip blocks the origin of the MCA. The resultingcessation of blood flow gives rise to subsequent brain infarction in theMCA territory (Longa, E. Z. et al., Reversible Middle Cerebral ArteryOcclusion Without Craniectomy in Rats, Stroke, 20, 84-91 (1989)). Thistechnique is used to study a temporary occlusion in which the MCA wasblocked for one hour. The filament is then removed allowing reperfusionto occur for 24 hours before the animal's brain was imaged usingT2-weighted scans in a 9.4T Bruker MRI scanner at the Case Center forImaging Research (FIG. 11). FIG. 11 shows a single slice from aT2-weighted MR image depicting the center of the brain showing cerebralcortex, hippocampus, thalamus, amygdala and hypothalamus for a “Normal”mouse (left panels) and a mouse which receives MCAo for one hourfollowed by 24 hours of reperfusion (right panels). Dashed lines markthe midline of the brain and show a large shift in the MCAo brain due tocerebral edema. Solid line highlights the region of infarct in the MCAobrain.

Survival—

Mice are treated with Compound 5 using a 2 mg/kg i.p. loading dose and 1mg/ml at 8 μl/h maintenance dose (delivered by an i.p. osmotic pump) ofCompound 5, or given saline (controls; n=17) using an identicalapproach. In this model, we observed a 29.4% improvement in overallsurvival at 24 h when animals are treated with Compound 5 (X²(1)=4.26;P<0.05).

Cerebral Edema—

Mice are given saline or treated with Compound 5 by multi-dosing at 5mg/kg i.p. every three hours (n=8 per treatment). This dosing regimen issufficient to maintain a plasma concentration of Compound 1>10 ng/ml forthe duration of the study. Ipsilateral and contralateral hemisphericvolume is measured from the T2-weighted MR images of mice 24 hourspost-icus. Relative change in hemispheric volume is calculated as apercent of the difference between ipsilateral brain volume (V_(i)) andcontralateral brain volume (V_(c)) relative to the contralateral brainvolume (Percent Change in Hemispheric BrainVolume=((V_(i)−V_(c))/V_(c))×100%.

Control animals show swelling in the ipsilateral hemisphere with arelative change in ipsilateral brain volume of 13.4%±1.9%, while animalsgiven Compound 5 show a 4.2±1.7% change (P=0.003, ±SEM, see FIG. 12).This represents a 3.2-fold reduction in brain swelling after MCAo.

Neurological Outcome—

In the same experiment as above, animals are scored for neurologicaloutcome on a simple 5 point scale described in Manley, G. T. et al.,Aquaporin-4 Deletion in Mice Reduces Brain Edema After Acute WaterIntoxication and Ischemic Stroke, Nature Medicine, 6, 159-163 (2000). Animprovement in neurological outcome is observed for animals givenCompound 5. Control animals have an average neurological score of2.77±0.66, while animals given Compound 5 have an average score of0.88±0.31 (FIG. 13, inset, P=0.025, n=9 per treatment). Animals givenCompound 5 did not progress into a state of severe paralysis or death.

The data from the MCAo stroke model together with the water toxicity(brain edema) model link the pharmacology of Compound 5/Compound 1 withimproved outcomes in stroke.

The invention claimed is:
 1. A method of treating or controllingcytotoxic cerebral edema comprising administering to a patient in needthereof an effective amount of

in free or pharmaceutically acceptable salt form.
 2. The method of claim1, wherein the cytotoxic cerebral edema is consequent to a trauma, astroke, a glioma, meningitis, acute mountain sickness, an epilepticseizure, an infection, a metabolic disorder, hypoxia, waterintoxication, hepatic failure, hepatic encephalopathy, or diabeticketoacidosis.
 3. The method of claim 1, wherein the cytotoxic cerebraledema is consequent to an abscess, eclampsia, Creutzfeldt-Jakob disease,or lupus cerebritis.
 4. The method of claim 1, wherein the cytotoxiccerebral edema is consequent to microgravity exposure, radiationexposure, or an invasive central nervous system procedure.
 5. The methodof claim 4, wherein the invasive central nervous system procedure isneurosurgery, an endovascular clot removal, a spinal tap, an aneurysmrepair, or deep brain stimulation.
 6. The method of claim 1, wherein thecytotoxic cerebral edema is consequent to a stroke.
 7. The method ofclaim 1, wherein the cytotoxic cerebral edema is consequent to a headtrauma.
 8. The method of claim 1, wherein the cytotoxic cerebral edemais consequent to an ischemic stroke.
 9. The method of claim 1, whereinthe cytotoxic cerebral edema is consequent to a traumatic brain injury.10. The method of claim 1, wherein the cytotoxic cerebral edema isconsequent to a middle cerebral artery stroke.
 11. The method of claim1, wherein the cytotoxic cerebral edema is consequent to hypoxia. 12.The method of claim 11, wherein the hypoxia is caused by cardiac arrestor other interruption of blood perfusion to the brain.
 13. The method ofclaim 11, wherein the hypoxia is caused by a stroke.
 14. The method ofclaim 1, wherein the patient has suffered a stroke or head injury within12 hours of commencing treatment.
 15. The method of claim 1, wherein thepatient has suffered a stroke or head injury within 6 hours ofcommencing treatment.
 16. The method of claim 1, wherein the patient hassuffered a stroke or head injury within 3 hours of commencing treatment.17. The method of claim 14, wherein the patient has suffered a strokewithin 12 hours of commencing treatment.
 18. The method of claim 15,wherein the patient has suffered a stroke within 6 hours of commencingtreatment.
 19. The method of claim 16, wherein the patient has suffereda stroke within 3 hours of commencing treatment.